HK1176287A - Methods for treatment of pain - Google Patents

Description

Method of treating pain

The present invention claims priority from provisional application No. 61/228,946 filed on 27.7.2009 in the united states, the disclosure of which is incorporated herein in its entirety.

The invention was accomplished in part with the help of NIH funding No. 1R43DE020816-01. The united states government has certain rights in this invention.

Background

Pain is sensation mediated by the excitation of certain brain structures. Pain is usually caused by activation of specialized nerve cells, nociceptors, when distributed in the skin or other surrounding tissues by mechanical, thermal, chemical or other noxious stimuli. Pain can also occur when peripheral or central nervous tissue becomes abnormally active in processes involving pain, such as a traumatic, ischemic or inflammatory pain event. Other causes of pain are: the process of a particular disease, metabolic disorder, muscle spasm, onset of a neurological event or syndrome.

Pain treatments of almost any type generally include one or more analgesic drugs, which are generally divided into three categories: mainly non-opioids, and compound analgesics, also known as adjuvants. Non-opioid analgesics include acetaminophen and non-steroidal anti-inflammatory drugs or NSAIDs. These drugs are effective in treating mild to moderate pain, but may have significant side effects such as acetaminophen-induced liver damage and NSAIDs-induced gastric ulcers.

Opioids, sometimes also referred to as "drugs", include natural substances such as opiates, opiate-derived substances such as morphine, and semi-synthetic and chemically synthetic substances such as fentanyl. Complex analgesics (Co-analgesic mediations) are narcotics generally directed to their indications, and are not used to relieve pain, but have an analgesic effect on certain pain conditions. An example of a compound analgesic is gabapentin (gaba transdermal pentin), which was originally indicated for the treatment of epilepsy, but also has a positive effect on the management of certain neuropathic pain.

Opioids are commonly used to relieve moderate to severe pain. However, their effects are limited because tolerance and dependence are generated in the course of normal treatment of chronic hepatitis b. Opioids such as morphine can be addictive and have significant and potentially fatal side effects such as respiratory depression, constipation, nausea, sensitivity to amplified pain, hormonal disturbances and changes in the immune system in addition to analgesia and mental changes.

Despite the wide range of available medications, pain continues to afflict millions of people in the united states alone, and remains a heavy burden on patients, healthcare, and businesses.

Disclosure of Invention

The methods and compositions disclosed herein for treating pain are by treatment with NOP agonists administered to the craniofacial mucosa, e.g., nasal administration of nociceptin (also known as nociceptin)/nociceptin (N/OFQ).

In one aspect, the invention provides a method of treating pain comprising administering to a craniofacial mucosa of an individual in need thereof an effective amount of an NOP agonist. In some embodiments, the treatment includes treatment of trigeminal neuralgia, somatic pain, neuropathic pain, post-operative pain, and muscular pain. In some embodiments, the treatment comprises acute pain and chronic pain. In some embodiments, the pain may be craniofacial pain or cephalic pain. In some embodiments, the craniofacial or cephalic pain is caused by a temporomandibular joint disorder (TMJ), migraine, or trigeminal neuralgia.

In some embodiments, the craniofacial mucosal administration comprises nasal administration, buccal administration, sublingual administration, or conjunctival administration. In a particular embodiment, the mucosal administration is intranasal administration. In some embodiments, the NOP agonist is administered less than two-thirds of the nasal cavity. In some embodiments, in particular, the NOP agonist is administered to less than two-thirds and more than one-third of the nasal cavity. In some embodiments, the NOP is administered through the conjunctiva or through other periocular tissues.

In particular embodiments, the NOP agonist is N/OFQ. In some embodiments, any of the methods further comprises administering to the subject in need thereof at least one additional active agent, wherein the additional active agent can be administered prior to, after, or simultaneously with the administration of the NOP agonist. In some embodiments, further, the method may comprise administering to the subject in need thereof at least 2 additional active agents, wherein the additional active agents may be administered prior to, after, or simultaneously with the administration of the NOP agonist.

In another aspect, the invention provides a method of treating pain comprising administering to an individual in need thereof craniofacial mucosal administration of an effective dose of an NOP agonist, wherein the administration results in a global analgesic effect. In some embodiments, the treatment comprises alleviation of pain. In some embodiments, the treatment comprises prevention of pain. In some embodiments, the craniofacial mucosal administration comprises nasal administration, buccal administration, sublingual administration, or conjunctival administration. In a particular embodiment, the mucosal administration is intranasal administration.

In one aspect, the invention provides a method of treating pain comprising administering intranasally to a subject in need thereof an effective amount of N/OFQ. In some embodiments, the dosage unit for N/OFQ administration is from about 0.2mg to about 5000 mg. In some embodiments, administration results in a reduction in VAS pain score (pain rating) of 30% or more.

In another aspect, the invention provides a method for treating or preventing migraine pain or treating migraine comprising administering to an individual in need thereof an effective amount of an NOP agonist, wherein the NOP agonist is administered via craniofacial mucosa, e.g., intranasally. In some embodiments, the treatment includes treating one or more symptoms associated with migraine headache, such as nausea, photophobia, and phonophobia (phonophobia). In some embodiments, the migraine is a migraine without aura, a migraine with aura, or a migraine with aura but without headache. In a particular embodiment, the method of treating migraine comprises nasal administration of an effective dose of N/OFQ to an individual in need thereof.

The invention also provides pharmaceutical compositions comprising an analgesic ingredient (e.g., a non-opioid analgesic peptide, a NOP agonist or N/OFQ), and methods of administering such pharmaceutical compositions via the craniofacial mucosa (e.g., intranasally, buccally, sublingually, or conjunctivally). And kits comprising the pharmaceutical compositions and optional administration devices. Further, the pharmaceutical composition further comprises one or more pharmaceutically acceptable excipients, adjuvants, diluents or stabilizers. In some embodiments, the pharmaceutical composition or the kit further comprises one or more additional activators, such as at least one additional analgesic component, a vasoconstrictor, at least one protease inhibitor and/or at least one absorption enhancer.

In another aspect, the invention also provides the use of an analgesic compound (e.g., a non-opioid analgesic peptide, an NOP agonist or N/OFQ), or a pharmaceutical composition comprising an analgesic compound (e.g., a non-opioid analgesic peptide, an NOP agonist or N/OFQ), in the manufacture of a medicament for the treatment of pain by administration to the craniofacial mucosa (e.g., intranasal administration). Also provided is the use of an analgesic compound (e.g., a non-opioid analgesic peptide, a NOP agonist or N/OFQ) for treatment of pain by craniofacial mucosal administration (e.g., intranasal administration).

Drawings

Figure 1 shows an a-delta experiment of cheek retraction latency in response to thermal stimuli: i.n. pain-producing hormone (0.2mg/kg, 1.0mg/kg and 5.0mg/kg) versus i.v. pain-producing hormone (5.0mg/kg) versus i.m. morphine (0.25mg/kg) versus i.m. saline. Data are presented as mean ± 1 SEM. (n is 6 rats/group)

Figure 2 shows a C-fiber experiment of cheek retraction latency in response to thermal stimuli: i.n. pain-producing hormone (0.2mg/kg, 1.0mg/kg and 5.0mg/kg) versus i.v. pain-producing hormone (5.0mg/kg) versus i.m. morphine (0.25mg/kg) versus i.m. saline. Data are presented as mean ± 1 SEM. (n is 6 rats/group)

FIG. 3 shows an A-delta experiment of paw withdrawal latency in response to thermal stimuli:

i.n. pain-causing hormone (0.2mg/kg, 5.0mg/kg) versus i.v. pain-causing hormone (5.0mg/kg) versus i.m. morphine (0.25mg/kg) versus i.m. normal saline. Data are presented as mean ± 1 SEM. (n is 6 rats/group)

FIG. 4 shows C-fiber experiments of paw withdrawal latency in response to thermal stimuli:

i.n. pain-causing hormone (0.2mg/kg, 5.0mg/kg) versus i.v. pain-causing hormone (5.0mg/kg) versus i.m. morphine (0.25mg/kg) versus i.m. normal saline. Data are presented as mean ± 1 SEM. (n is 6 rats/group)

Figure 5 shows a C-fiber experiment of cheek retraction latency in response to thermal stimuli: the pain-producing hormone antagonist (10.0mg/kg) plus i.n. pain-producing hormone (5.0mg/kg) was injected subcutaneously versus the normal saline plus pain-producing hormone (5.0mg/kg) injected subcutaneously. Data are presented as mean ± 1 SEM. (n is 3 rats/group)

Figure 6 shows a C-fiber experiment of cheek retraction latency in response to thermal stimuli: the ratio of the antagonist of the analgesic (10.0mg/kg) to the i.n-analgesic (5.0mg/kg) was compared to the ratio of the physiological saline + the analgesic (5.0 mg/kg). Data are presented as mean ± 1 SEM. (n is 3 rats/group)

FIG. 7 shows A-delta experiments on paw withdrawal latency in response to thermal stimuli:

the ratio of the antagonist of the analgesic (10.0mg/kg) injected subcutaneously to the i.n. analgesic (5.0mg/kg) was compared with the normal saline plus the analgesic (5.0mg/kg) injected subcutaneously. Data are presented as mean ± 1 SEM. (n is 3 rats/group)

FIG. 8 shows C-fiber experiments of paw withdrawal latency in response to thermal stimuli:

the ratio of the antagonist of the analgesic (10.0mg/kg) to the i.n-analgesic (5.0mg/kg) was compared to the ratio of the physiological saline + the analgesic (5.0 mg/kg). Data are presented as mean ± 1 SEM. (n is 3 rats/group)

Figure 9 shows the comparison of analgesic activity of nasally administered N/OFQ and morphine in response to formalin injected labial-rostral male rats. The face Time (Time period grooming) exceeded 45 minutes and was divided into groups every 3 minutes. The nasal administration has the following component concentrations: morphine 3.0 mg/mouse and N/OFQ 0.3 mg/mouse. The area of the curve (AUC) for each ingredient or Vehicle (Vehicle) was determined by a similar trapezodial method. The% reduction is [ (vehicle AUC treatment AUC)/vehicle AUC) ] x 100% data are presented as mean ± 1 SEM. (n-6-8 rats/group).

FIG. 10 shows dose responses of post-operative nasal administration of 1, 3, 5 and 10mg/kg N/OFQ to punctate allodynia caused by incisions in the mouse paw. Data are presented as mean ± 1 SEM. (n-6-13 rats/group). -p < 0.05 &.

FIGS. 11 and 12 show the anti-allodynia (anti-allodynic) activity of N/OFQ administered to male rats on static allodynia following surgical incision of the plantar surface of the hind limb. N/OFQ was administered nasally at a concentration of 5.0mg/kg 24 hours after incision (FIG. 11) or intravenously at a concentration of 5.0mg/kg 24 hours after incision (FIG. 12). The 50% threshold for withdrawal was determined from top to bottom using von Frey films every 30 minutes after dosing. Data are presented as mean ± 1 SEM. (n-5-6 rats/group). P < 0.05 compared to vehicle.

Figure 13 shows the anti-allodynic activity of N/OFQ administration to male rats after cheek surgery incision for static allodynia. N/OFQ was administered nasally at a concentration of 5.0mg/kg 24 hours after incision. The 50% threshold for withdrawal was determined from top to bottom using von Frey films every 30 minutes after dosing. Data are presented as mean ± 1 SEM. (n-4-5 rats/group). P < 0.05 compared to vehicle.

FIG. 14 shows dose responses of 1, 3, 5 and 10mg/kg N/OFQ administered nasally after surgery for punctate allodynia caused by incisions in the mouse cheek. Data are presented as mean ± 1 SEM. (n-5-13 rats/group). Compared with the postoperative value, # -p < 0.05& # # p < 0.01.

FIG. 15 shows the anti-allodynic activity of static punctate allodynia N/OFQ administration in a Spared Nerve Injury (SNI) experiment in male rats. N/OFQ was administered intranasally at a concentration of 5.0mg/kg for 21 days after surgery. The 50% threshold for withdrawal was determined from top to bottom using von Frey films every 30 minutes after dosing. Data are presented as mean ± 1 SEM. (n-4-6 rats/group).

Figure 16 shows the intranasal administration of N/OFQ (5.0mg/kg), the intranasal administration of morphine (4.0mg/kg), or the passage of morphine intraperitoneally (3.0mg/kg) through the intestinal tract (transit), expressed as% of vehicle (vehicle). The ingredients were administered 20-30 minutes prior to the charcoal meal (2 mL/rat). Rats were sacrificed 30 minutes after the meal. transit% is multiplied by 100% by the calculated ratio of the distance traveled by the charcoal label divided by the full length of the small intestine. Data are presented as mean ± 1 SEM. (n-8 rats/group). Provided are pharmaceutical compositions, methods and others for treating pain by administration of analgesic compounds, such as non-opioid analgesic peptides, polypeptides, or proteins (e.g., nociceptin/orphyrin (N/OFQ) or other non-opioid molecules, via craniofacial mucosa.

Definition of

Although analgesia (analgesia) is the elimination of pain in the strict sense, as used herein,

"analgesia" (analgesia) refers to the reduction in pain perception in an individual.

An "analgesic" (Analgesia agent), "analgesic" (analgesic agent) or "analgesic" (analgesic) refers to any biomolecule, drug or active agent that reduces or prevents pain. "Acute pain" (Acute pain) refers to sudden pain (as opposed to chronic pain) of a limited duration, caused by a specific cause (injury, infection, inflammation, etc.). "Chronic pain" (Chronicpain) refers to pain in a persistent state. Chronic pain is often associated with a long-term incurable or refractory clinical condition or disease. "post-operative pain" (Procedural pain) refers to pain arising from medical, dental or other procedures in which severe trauma may be planned or involved. "headache disorder" includes migraine, tension pain, combined headache, trigeminal neuralgia, secondary headache, and miscellaneous headache.

"migraine" includes migraine headaches, migraine without aura, migraine with aura, and migraine with aura but without headache.

"systemic side effects", including, but not limited to, cardiovascular disease, including peripheral vasodilation and baroreceptor inhibition; skin disorders, including skin itch (pruritus), flushing and red eyes; gastrointestinal disorders including nausea, vomiting, and the like, decreased gastrointestinal motility, decreased bile secretion, delayed secretion of pancreatic and intestinal secretions and digestive food, decreased motility in the large intestine resulting in constipation, epigastric discomfort or biliary colic, and respiratory disorders including respiratory distress; urinary problems include urgency, difficulty in urination; and heavy peripheral limbs.

"side effects of the central nervous system" or "side effects of the central nervous system" (CNS sideeffects) include, but are not limited to, anesthesia, excitement, lethargy, apathy, psychosis, confusion, mood changes, decreased body temperature, relaxation of feelings, irritability (anxiety, depression, or characteristic of restless mood states), nausea, and vomiting (caused by direct stimulation of bulbar chemoreceptors).

By "systemic analgesic effect" is meant the effect on any part of the body. Systemic analgesic effects may be achieved with or without systemic circulation of the active agent. For example, a systemic analgesic effect may be achieved by nasal administration of nociceptin, which may not result in significant systemic circulation of N/OFQ peptide.

"craniofacial mucosal administration" means through the mucosal surfaces of the nose, nasal passages, nasal cavities, oral mucosa including gums (gums), oral floor, lips, tongue; and ocular mucosal surfaces or surroundings, including conjunctiva, lacrimal gland, lacrimal duct, upper or lower eyelids and ocular mucosa, and the eye.

By "nasal administration" or "intranasal administration" is meant administration to the nose, nasal passages or nasal cavities by spraying, dripping, dusting, gelling, film, inhalation or other means.

The "nasal cavity disadvantaged region" refers to the part of the nasal cavity where the turbinate bones protrude, which is a region of the nasal cavity where trigeminal innervation is evident. The "nasal dominance area" is defined as the third and cribriform plate area, where the olfactory innervation is located.

Nociceptin (Nociceptin)/Nociceptin (N/OFQ) is a 17 amino acid peptide, with a molecular weight of 1809, belonging to the family of opiate peptides. The human N/OFQ amino acid sequence is:

it binds to a G protein-coupled receptor commonly known as ORL1, now formally designated the NOP receptor. Although the NOP receptor is a protein of the opioid receptor family, opioids do not bind to the NOP receptor as tightly as possible; the binding of N/OFQ to opioid receptors is also not tight.

Thus, N/OFQ is considered a non-opioid peptide. When N/OFQ was first discovered and tested in mice by the lateral ventricle (ICV) route of administration, it was found that mice had reduced tail flick and reduced hot plate latency. This indicates that, unlike opioid polypeptides, N/OFQ is pain responsive, rather than pain free. Further studies by several groups of subjects indicated that i.c.v.n/OFQ blocked opioid analgesics from being mediated through each opioid receptor.

However, intrathecal injection of (i.t)/OFQ potentiates the analgesic effect of morphine or analgesia is achieved by itself. This indicates that the site of administration of N/OFQ is important for its ultimate utility. When the first NOP receptor antagonist was designed, it was tested by i.c.v. administration and found to have analgesic effect.

Like other members of the opioid receptor family, the NOP receptors are Gi/O coupled. Without being bound by any particular theory, activated NOP receptors may lead to a decrease in intracellular cAMP levels, which also inhibit the potassium channel of G protein-coupled receptors (GIRK channel).

This may lead to a decrease in the polarization and cellular activity of the cell in which the receptor is located. N/OFQ may act in the brain by reducing neuronal activity, which may lead to attenuation of endogenous or exogenous opioid activity, thereby relieving inhibition of the pain pathway. In the spinal cord, cellular localization may act like analgesics, similar to opioids, but the mechanism of action may differ.

The NOP receptors and N/OFQ are found in many areas throughout the brain and thus have many central nervous system behaviors. One region with a high number of NOP receptors is the trigeminal ganglion. This is also the brain region that mediates craniofacial and cephalic pain due to a variety of conditions including temporomandibular joint disorder (TMJ), migraine, trigeminal neuralgia and the like.

Intranasal administration of N/OFQ may focus on the trigeminal ganglia and reduce the activity of these neurons. Although the inhibitory effect of these neurons is not known without testing, one possibility is regional analgesia.

In addition, it is not known whether N/OFQ will reach concentrations in other parts of the brain high enough to suppress the analgesic response (as seen in i.c.v. administration of N/OFQ). Given that N/OFQ has different functions, hyperalgesia is present in some areas of the CNS and in other areas of the CNS and in nerve endings, all the results of N/OFQ after intranasal administration are unpredictable.

In the thermal pain rat model, N/OFQ intranasal administration showed dose-dependent analgesic effects in the face of A-delta and C-fiber pain threshold tests in thermally induced pain, while N/OFQ showed low or no effect when administered intravenously.

This indicates that the analgesic effect of N/OFQ administered nasally is not a result of systemic blood circulation. A more surprising result was observed that heat caused pain in the rat hind paw, and that nasal administration of N/OFQ showed a dose-dependent analgesic effect on pain induced by heat in the rat hind paw in both A-delta and C-fiber pain threshold experiments (FIGS. 3 and 4). Experiments with intravenous orphanin in the paw showed little or no effect.

Thus, when administered intranasally, N/OFQ shows an analgesic effect on pain not only in one area (e.g. on the face), but also systemically, whereas systemic administration of the same dose (e.g. i.v. administration) shows little or no analgesic effect at all. In fact, i.n. orphanin produces a systemic analgesic effect, whereas systemic administration has no effect, possibly implying that i.n/OFQ may activate a down-regulated pain-regulating system. Thus, the present invention provides a method for treating pain using intranasal N/OFQ, wherein the result of intranasal administration is a systemic analgesic effect.

Also highly useful in the methods of the invention are agonists of the NOP receptor or NOP agonists, including both peptidic and non-peptidic compounds. Examples of NOP agonist Peptides include, but are not limited to, N/OFQ, shortened N/OFQ analogs (Reinscheid, R.K.et al, J.biol.chem. (1996) 271: 14163-. Small molecule NOP agonists include, for example, hexahydrospiro [ pipe idine-4, 1' -pyrrolo [3, 4-c ] pyrroles ] (Kolczewski, S.et al, J.Med.chem. (2003) 46: 255-264) and other non-peptide NOP agonists. Although nasal, conjunctival, oral delivery of analgesic molecules for the treatment of head and facial pain has been proposed (us 20070093420, us 20070054843), to date there is no definitive evidence of the efficacy of nasal, conjunctival, oral delivery of NOP agonists for the treatment of pain.

Many peptides are considered analgesics. They are administered by various routes to provide local, regional and/or systemic analgesic effects. There is a class of analgesic peptides, known as "opioid peptides", which have an opioid receptor binding moiety that binds with high affinity to opioid receptors. Opioid peptide is a naturally occurring endogenous peptide fragment, analog or derivative. The opioid peptide may also be a non-endogenous peptide fragment, analog or derivative. Examples of opioid peptides are endorphins, enkephalins, dynorphins, dermorphins (dermorphins), picorphins (dermenkephalins), morphine-sensitive receptors (morphine), endomorphins, dalargin (dalargin), and other synthetic mu, delta and kappa agonist peptides.

As used herein, "non-opioid analgesic peptide" refers to an analgesic peptide that reduces or prevents pain by a mechanism other than its high affinity for opioid receptors or a peptide that has a lower affinity for opioid receptors than is generally considered an opioid or opioid drug molecule. For example, an analgesic peptide having a low affinity for the mu-, delta-and kappa opioid receptors (e.g., less than about 1 micron or less than about 10 microns) is considered a non-opioid analgesic peptide.

Unless otherwise indicated, binding affinity herein is the dissociation constant (KD) in units of one molar concentration (m), with half of the binding site of a particular protein being the concentration of the corresponding ligand when occupied, i.e., the concentration of ligand at which the receptor binds to the ligand, equal to the concentration at which the ligand does not bind to the receptor. The smaller the dissociation constant, the tighter the binding of the ligand and the higher the affinity between the ligand and the protein.

For example, a ligand with a nM dissociation constant binds more tightly to a particular protein than a ligand with a μ M dissociation constant. When a ligand binds to a receptor with an affinity of less than 1. mu.M, the dissociation constant for receptor-ligand binding is higher than 1. mu.M.

Examples of non-opioid analgesic peptides for use in the methods of the present invention include, but are not limited to, those selected from the group consisting of optic juxtins (hydocretens)/orexins (orexins), calcitonin (calcein), octreotide (octreotide), somatostatin (somatotatin), vasopressin (vasopressin), galanins, lipotropin C fragments (C-fragments of lipotropin) and Ac-rfwink-NH2, omega-conotoxin GV1A (omega-conoxin GV1A), omega-conotoxin MVIIA, peptide antagonists of the pain-peptide neurotransmitter receptors CGRP 1and CGRP2, such as CGRP 8-37 and CGRP 28-3; analgesic peptide neurotransmitter receptors NK1 including N-acetyl trptophan, D-Pro9- [ Spiro-y-lactam]-Leu 10, Trp 11-Physalaemin (1-11), Tyr-D-Phe-Phe-D-His-Leu-Met-NH2 (Sende) and splatide II. Analgesic peptide neurotransmitter receptors NK2, including PhCO-Ala-Ala-D-Trp-Phe-D-Pro-Pro-Nle-NH₂(GR98400)，[Tyr5，D-Trp6，8，9，Lys 10]-NKA (4-10) (MEN10376) and derivatives thereof. The pain peptide neurotransmitter receptors VPAC2, VPAC1and PAC1 include VIP (6-28), Ac His (1) [ D-Phe (2), K (15), R (16), L (27)]VIP(3-7)/GRF(8-27)。

U.S. patent No. 7220725 discloses the use of pyroglutamic acid tripeptide and tetrapeptides in the form of ointments, creams or salves for the treatment of pain. The pyroglutamic acid tripeptides (tripeptides) and tetrapeptides (tetrapeptides) disclosed in U.S. patent No. 7220725 may also be used for the treatment of pain by intranasal administration in the present invention. Proteins that provide analgesic effects may also be used in the methods of the invention, such as Brain Derived Neurotrophic Factor (BDNF), Nerve Growth Factor (NGF), neurotrophic factor-3 (NT-3), botulinum toxin, anti-inflammatory cytokines (e.g., interleukin 4, interleukin-10, and interleukin-13)

The compounds used in the methods of the invention can be evaluated by appropriate assays in the art, such as human clinical studies and animal model trials for the treatment of pain. In some embodiments, craniofacial mucosal administration of the compounds used in the methods of the invention exhibits a systemic analgesic effect, such as by intranasal administration. In some embodiments, the compounds used in the methods of the invention exhibit greater analgesic effect by nasal administration than by intravenous administration. In some embodiments, the compounds exhibit little or no analgesic effect when administered by intravenous injection. In some implementations, craniofacial mucosal administration of the compound in the methods of the invention can result in significant systemic blood circulation of the compound. In some embodiments, craniofacial mucosal administration of the compounds in the methods of the invention can result in some blood circulation of the composition. In some embodiments, craniofacial mucosal administration of the compounds does not cause significant central nervous system side effects.

Pain (due to cold or dampness)

Pain is an unpleasant sensory and emotional experience associated with actual or potential tissue damage, which may be as described by (http:// www.iasp-pain. org /), as classified in Table 1 according to recommendations in the International society for pain research.

TABLE 1

Pain treatable using the methods of the invention includes any of the pain described herein, e.g., any combination of pain as described in table 1.

In certain aspects, the invention provides methods of treating pain by administering an analgesic component (e.g., a non-opioid analgesic peptide, a NOP agonist or N/OFQ) to the craniofacial mucosa (e.g., intranasally, buccally, sublingually or conjunctivally). In some embodiments, the pain is somatic pain. In some embodiments, the pain is a superficial somatic pain. In some embodiments, the pain is a deep somatic pain. In some embodiments, the pain is a musculoskeletal pain. In some embodiments, the pain is a visceral pain. In some embodiments, the pain is a neuropathic pain. In some embodiments, the pain is a head pain or a craniofacial pain. In some embodiments, the pain is perioperative/perioperative pain. In some embodiments, the present invention provides a safe and effective treatment that does not cause severe sedation, respiratory depression, and/or addiction distress.

In some embodiments, the present invention provides methods of treating acute pain by administering an analgesic compound (e.g., a peptide other than an opioid analgesic, a NOP agonist, or/OFQ) to the craniofacial mucosa (e.g., nasally, buccally, sublingually, or conjunctivally). Examples of acute pain include, but are not limited to, traumatic pain, procedural (surgical, stomatological, dermatological, etc.) pain, wound care pain, headache (migraine, cluster, etc.), musculoskeletal/spinal pain (e.g., back pain), ear pain, dental pain, ischemic (e.g., heart) pain, acute glaucoma, gastrointestinal disorders (cholecystitis, volvulus, appendicitis, etc.), urological disorders (calculus, volvulus, etc.), spasticity, infection, inflammation-induced pain (gout, lupus erythematosus, etc.), toxins (insects, animals, etc.), angina, menstrual cycle, and labor pain.

In some embodiments, the non-opioid analgesic peptide, NOP agonist or N/OFQ inhibits exercise-related acute sting. This type is of great interest in perioperative settings if the pain is opioid poor coverage. In some embodiments, intranasal delivery of a non-opioid analgesic peptide, NOP agonist or N/OFQ treats acute procedural pain such as pain experienced during surgery (e.g., a patient undergoing major surgery such as hip and knee replacement) or post-surgery.

In some embodiments, the present invention provides methods of treating chronic pain by craniofacial mucosal administration (e.g., intranasal, buccal, sublingual, or conjunctival) of an analgesic compound (e.g., a peptide other than an opioid analgesic, a NOP agonist, or/OFQ). Examples of chronic pain include, but are not limited to, fibromyalgia, arthritic pain, cancer pain, musculoskeletal/spinal pain (including back pain), temporomandibular joint disorder (TMJD or TMJ), trigeminal neuralgia, chronic headache, complex regional pain syndromes (types I and II), pain associated with neurological disease (MS, diabetic neuropathy, etc.), neuroma, pelvic inflammatory disease, endometriosis, postherpetic neuralgia and shingles, infections associated with chronic neuropathy (e.g., HIV), neuropathic pain induced by chemotherapy, neuropathic pain induced by surgery, neuropathic pain induced by trauma, vulvodynia, atypical cranial facial pain, cervical spondylotic radiculopathy/sciatica, phantom limb (phantom), dental pain, burning mouth syndrome.

Migraine and headache

Various medical conditions that can cause headache pain include, but are not limited to, primary headaches such as migraine, cluster headaches, tension or stress headaches, chronic daily headaches, secondary headaches caused by specific conditions such as secondary headaches caused by tumors and infectious diseases, toxin intake or over-drinking, and trigeminal neuralgia. As described herein, a common type of primary vascular headache is migraine, with cluster headaches being less common but also debilitating. Tension or "stress" type headaches are considered to be the most common headaches of all types, considering the number of affected individuals is the greatest. The characteristics of these headaches or headache disorders are described in HIS and summarized in table 2.

TABLE 2

Headache and trigeminal neuralgia are often not effectively treated by the current medicines, and new methods for relieving pain are urgently needed. Thus, in some aspects of the invention, methods are provided for treating headache or trigeminal neuralgia by administering an effective amount of a NOP agonist, wherein the administration results in analgesia at the junction or head region. NOP agonists can be administered to patients with headache, including, but not limited to, migraine, cluster headache, tension headache, secondary forms of headache and trigeminal neuralgia.

In certain aspects, the invention provides methods of treating migraine or other pain comprising administering to an individual in need thereof an effective amount of an analgesic compound (e.g., a non-opioid analgesic peptide, a NOP agonist, or N/OFQ) via craniofacial mucosal administration (e.g., nasal, buccal, sublingual, or conjunctival administration). In some embodiments, the invention provides methods of treating migraine comprising nasally administering to an individual in need thereof an effective dose of N/OFQ.

In some embodiments, the migraine is migraine without aura, migraine with aura, or migraine with aura but without headache, basilar artery type migraine, familial hemiplegic migraine, sporadic hemiplegic migraine, abdominal type migraine, non-headache migraine (aphtalgic migraine) or menstrual migraine. In some embodiments, the treatment comprises alleviating or preventing one or more symptoms associated with migraine. In some embodiments, the symptom is a prodrome stage symptom, a premonitory stage symptom, a pain stage symptom, or a late stage symptom. In some embodiments, the treatment comprises alleviating or preventing one or more of the symptoms of nausea, photophobia, and phonophobia.

Method of producing a composite material

In one aspect, the invention provides a method of treating pain in an individual, comprising: administering to the individual via the craniofacial mucosa an effective amount of an analgesic compound or a pharmaceutical composition comprising an analgesic compound. In certain aspects, analgesic compounds in the methods of the invention include, but are not limited to, peptides, amino acids, polypeptides or small molecule compounds having analgesic activity. In some embodiments, the analgesic component is a non-opioid analgesic peptide.

In some embodiments, the analgesic component is an analgesic peptide that binds to opioid receptors by a mechanism other than high affinity to reduce or prevent pain, and in some embodiments, is a peptide that has a lower affinity for opioid receptors than drug molecules that are generally considered opioids or opioids. In some embodiments, the analgesic component is an analgesic peptide having an affinity for mu-, delta-and kappa opioid receptors of less than 10mM, less than 1mM, less than 0.1mM, less than 10. mu.M, or less than 1. mu.M (e.g., less than about 1 micron or less than about 10 microns).

In some embodiments, the analgesic component is a non-opioid analgesic peptide selected from the group consisting of: hypocretinins/orexin, calcitonin, octreotide, somatostatin, vasopressin, galanin, lipotropin C fragment and Ac-rfwink-NH₂Omega-conotoxin GV1A, omega-conotoxin MVIIA, peptide antagonists of the pain peptide neurotransmitter receptors CGRP 1and CGRP2, such as CGRP 8-37 and CGRP 28-3; analgesic peptide neurotransmitter receptors NK1 including N-acetyl trptophan, D-Pro9- [ Spiro-y-lactam]-Leu 10，Trp 11-Physalaemin(1-11)，Tyr-D-Phe-Phe-D-His-Leu-Met-NH₂(Sende) and splantide II. Analgesic peptide neurotransmitter receptors NK2, including PhCO-Ala-Ala-D-Trp-Phe-D-Pro-Pro-Nle-NH₂(GR98400)，[Tyr 5，D-Trp6，8，9，Lys 10]-NKA (4-10) (MEN10376) and derivatives thereof. The pain peptide neurotransmitter receptors VPAC2, VPAC1and PAC1 include VIP (6-28), Ac His (1) [ D-Phe (2), K (15), R (16), L (27)]VIP (3-7)/GRF(8-27)。

In some embodiments, the analgesic component is the pyroglutamic acid tripeptides and tetrapeptides disclosed in U.S. patent No. 7220725. In some embodiments, the analgesic component is a polypeptide, such as brain-derived neurotrophic factor (BDNF), Nerve Growth Factor (NGF), neurotrophic factor-3 (NT-3), botulinum toxin, an anti-inflammatory cytokine (e.g., interleukin 4, interleukin-10, and interleukin-13.)

In some implementations, the analgesic component is an NOP receptor agonist. In some embodiments, the analgesic component is an NOP agonist peptide, including N/OFQ, shortened N/OFQ analogs (Reinscheid, R.K.et al, J.biol.chem. (1996) 271: 14163-. In some embodiments, the analgesic compound is a non-peptide NOP agonist such as hexahydrospiro [ piperidine-4, 1' -pyrrolo [3, 4-c ] pyrroles ] and other small molecule NOP agonists as described herein. In a particular embodiment the analgesic component is N/OFQ (orphanin).

In some embodiments, a method of treating pain is provided comprising administering to a subject in need thereof an effective amount of a non-opioid analgesic peptide (e.g., N/OPQ) through the craniofacial mucosa. Craniofacial mucosal administration includes, but is not limited to, intranasal administration, buccal administration, sublingual administration, and conjunctival administration. In some embodiments, prophylactic mucosal administration of a non-opioid analgesic peptide can prevent or delay the onset of pain. In some embodiments, mucosal administration of the non-opioid analgesic peptide reduces or alleviates severe pain.

In some embodiments, the pain is somatic pain. In some embodiments, the pain is a superficial somatic pain. In some embodiments, the pain is a deep somatic pain. In some embodiments, the pain is a musculoskeletal pain. In some embodiments, the pain is a visceral pain. In some embodiments, the pain is a neuropathic pain.

In some embodiments, the pain is a chronic pain of the invention. In some embodiments, the pain is an acute pain according to the invention. In some embodiments, the pain is pain associated with migraine, cluster headache, tension headache, secondary headache, or trigeminal neuralgia.

In some embodiments, the pain is pain caused by TMJ, migraine or trigeminal neuralgia. In some embodiments, the pain is acute sharp pain. In some embodiments, the pain is neuropathic pain resulting from surgery associated with a nerve injury.

In some embodiments, the method comprises administering to the individual via craniofacial mucosal administration an analgesic component (e.g., a non-opioid analgesic peptide, a NOP agonist or N/OFQ) or a pharmaceutical composition comprising an analgesic component (e.g., a non-opioid analgesic peptide, a NOP agonist or N/OFQ) to treat migraine-related pain, cluster headache or trigeminal neuralgia.

In some embodiments, the method comprises treating migraine-associated pain by abortive treatment (abortive treatment) by administering to an individual experiencing migraine or other headache, via the craniofacial mucosal route, an analgesic composition or a pharmaceutical composition comprising an analgesic composition. In some embodiments, the method comprises prophylactically treating (pharmacological treatment) migraine headache-associated pain comprising preventing the onset of migraine headache pain by administering to an individual via the craniofacial mucosal route to an analgesic agent or a pharmaceutical composition comprising an analgesic agent.

In some embodiments, the method comprises prophylactically treating migraine-associated pain comprising administering to an individual experiencing a prior precursor to migraine headache associated with migraine headache via craniofacial mucosal route to an analgesic component or a pharmaceutical composition comprising an analgesic component. In some embodiments, the method comprises prophylactically treating cluster headache pain comprising administering to an individual who has begun but has not begun a cluster continuation of headache via craniofacial mucosal route administration of an analgesic composition or a pharmaceutical composition comprising an analgesic composition.

In some embodiments, the method comprises prophylactically treating trigeminal neuralgia comprising administering to an individual having an onset of trigeminal neuralgia but not having a consecutive onset, via the craniofacial mucosal route to the analgesic composition or to a pharmaceutical composition comprising the analgesic composition.

In one aspect, the invention provides a method of treating pain comprising administering to a craniofacial mucosa of a subject in need thereof an effective amount of an analgesic component (e.g., a peptide other than an opioid analgesic, a NOP agonist, or/OFQ), wherein the result of the administration is a systemic analgesic effect.

In experiments investigating the analgesic effect of N/OFQ peptide when administered intranasally in animals, we found that N/OFQ had a longer latency period for cheek retraction (shown in both A-delta and C-fiber analgesic experiments) after intranasal administration than after administration of control saline, but no significant analgesic effect was found with the same dose of N/OFQ administered by injection.

In contrast to saline administration, the same analgesic effect was observed in paw withdrawal experiments when N/OFQ was administered intranasally, but this was not observed in the case of injection.

In some embodiments of the invention, an analgesic peptide, such as N/OFQ, is administered by nasal administration to achieve systemic analgesic effects. In some embodiments, craniofacial mucosal administration (e.g., intranasal administration) of an analgesic composition (e.g., N/OFQ) results in a greater analgesic effect than administration of the composition by injection. In some embodiments, craniofacial mucosal administration of the analgesic composition does not cause significant systemic side effects. In some embodiments, when a non-opioid analgesic peptide such as N/OFQ is administered intranasally, a substantial portion of the non-opioid analgesic peptide (e.g., N/OFQ) reaches the brain and is delivered to the brain without entering systemic blood circulation.

In one aspect, the invention provides a method of treating pain in an individual comprising administering to the individual an analgesic component (e.g., a non-ap analgesic peptide such as N/OFQ or other NOP agonist) to a mucosal tissue or to an oral epithelial tissue, to the nasal cavity, to the inside or periphery of the eye, or to the skin. Oral mucosal tissue includes, but is not limited to including the gums (gums), the floor of the mouth, lips, tongue, or combinations thereof.

The method includes administering an analgesic composition to the conjunctiva or other mucosal tissue surrounding the eye. Such tissues or epithelial tissues include, but are not limited to, conjunctiva, lacrimal gland, lacrimal duct, upper or lower eyelids and ocular mucosa, eye and combinations thereof. One component or pharmaceutical composition administered to the conjunctiva but not completely absorbed by the conjunctiva through the conjunctival mucosa may pass through the nasal tube into the nasal cavity where it may be absorbed by the mucosal tissue surrounding the nasal cavity.

In some embodiments, the analgesic component is a non-opioid peptide. In some embodiments, the analgesic component is a peptide or a non-opioid NOP agonist. In a particular embodiment, the analgesic component is an N/OFQ peptide. The analgesic component may be administered to mucosal tissue surrounding the nasal cavity. Suitable areas include, but are not limited to, less than two-thirds or more than one-third of the nasal cavity, or the entire nasal cavity.

In some embodiments, an NOP agonist (e.g., an NOP analgesic peptide, e.g., N/OFQ) is administered to more than one-third of the nasal cavity. In some embodiments, an NOP agonist (e.g., an NOP analgesic peptide, such as N/OFQ) is administered to less than two-thirds of the nasal cavity. In particular, in some embodiments, NOP agonists (e.g., NOP analgesic peptides, such as N/OFQ) are administered to up to two-thirds and one-third of the nasal cavity.

Intranasal administration has been the subject of research and development for many years, although drug delivery systems that can effectively deliver materials have been designed for only a decade. (Sayani and Chien, clinical reviews in Therapeutic Drug Systems 1996, 13: 85-184.) intranasal delivery has many advantageous functions, including high bioavailability, kinetics of rapid absorption and avoidance of the first pass effect (first-pass effect) of the liver.

The use of nasal administration provides a convenient, rapid, non-invasive mode of administration, considering patient compliance and ease of use. In some aspects, nasal administration may allow delivery of analgesic peptides such as N/OFQ into the nasal cavity, and in other aspects, nasal administration may allow targeted delivery to the nose and/or cranial nerves of the brain. Without wishing to be bound by any particular theory, nasal administration of an analgesic peptide, such as N/OFQ, may be directed to the olfactory or trigeminal nervous systems, or both.

As with oral and intravenous injections, intranasal delivery of peptides suffers from malabsorption and extensive hydrolysis if the effect of the administration of the peptide requires access to the systemic blood circulation. Enzyme inhibitors and absorption enhancers are used to increase the extent of peptide absorption. Leucine enkephalin, an opioid peptide, has shown very limited analgesic effects when tested in mice with acetate motility (visceral pain model) by nasal administration, while leucine enkephalin cocktail enzyme inhibitors and absorption enhancers have shown high analgesic effects when administered intranasally (Gawk h.s.et al, Journal of Pharmacy and Pharmacology 2003, 55: 1207. sup.1212), achieving enhanced systemic absorption and targeted delivery to cranial nerves and/or brain.

Oral, buccal or sublingual routes of delivery are convenient options for drug delivery because they are easy to use and non-invasive. The advantages include: (ii) in the oral cavity, the proteolytic activity in the oral cavity is low compared to other routes, thereby avoiding the problem of degradation of polypeptides and protein drugs by enzymes, and (ii) avoiding the first pass effect over the liver. Drug delivery to the mucosal tissue or conjunctiva surrounding the eye is another convenient option for non-invasive administration.

In certain aspects, the methods of the invention treat pain, such as headache pain or trigeminal neuralgia in an individual, by administering an effective dose of an analgesic peptide, such as N/OFQ, to the conjunctiva or other mucosal tissue surrounding the eye. Transdermal administration or administration of skin treatment agents to the skin has become a well established technique for the past 20 years. Transdermal drug delivery controls the release of the drug to the patient, is easy to use, convenient, painless, and provides for multiple days of dosing, which often results in increased patient compliance. These methods may include administering an analgesic component (e.g., a non-opioid analgesic peptide, NOP agonist or N/OFQ) or a pharmaceutical composition containing an analgesic component (e.g., a non-opioid analgesic peptide, NOP agonist or N/OFQ) to the skin of the face, head or body.

NOP agonists, can be administered to the skin of the face, scalp or temporal region. Suitable facial skin includes skin of the chin, upper lip, lower lip, forehead, nose, cheek, around the eyes, upper eyelid, lower eyelid, or combinations thereof. Suitable scalp skin includes the anterior scalp, the entire temporal scalp, the lateral portion of the scalp, or combinations thereof. The skin of the temporal region is suitable, including the entire temporal region and combinations thereof.

Intradermal injection is defined as the administration of a therapeutic drug within or between layers of the skin. In contrast, subcutaneous injection is defined as below the initial layer of skin. The administration of therapeutic agents by intradermal or subcutaneous injection is a common practice by those skilled in the art.

In some embodiments, administration of an analgesic component (e.g., a non-opioid analgesic peptide, NOP agonist or N/OFQ) or a pharmaceutical composition containing an analgesic component (e.g., a non-opioid analgesic peptide, NOP agonist or N/OFQ) via craniofacial mucosal administration results in the prevention or alleviation of pain without causing paralysis as compared to the strong sedative effect of local anesthetics or narcotics. The targeted delivery may reduce the amount of analgesic component administered to an individual to achieve analgesic effect and may reduce potential adverse central nervous system effects or systemic side effects.

In one aspect, the present invention provides a method of treating migraine in an individual comprising administering to the craniofacial mucosa an analgesic component (e.g., a non-opioid analgesic peptide, a NOP agonist or N/OFQ) or a pharmaceutical composition comprising an analgesic component (e.g., a non-opioid analgesic peptide, a NOP agonist or N/OFQ). In some embodiments, the craniofacial mucosal route is a nasal, oculoconjunctival, sublingual, or buccal route. In some embodiments, the method comprises treatment of migraine.

In some embodiments, the method comprises treatment of migraine without aura. In some embodiments, the method comprises treatment with a migraine headache. In some embodiments, the method comprises treatment of migraine with aura but without headache. In some embodiments, the method comprises treatment of basal-type migraine. In some embodiments, the method comprises treatment of familial hemiplegic migraine or sporadic hemiplegic migraine. In some embodiments, the method comprises treatment of abdominal migraine. In some embodiments, the method comprises treatment of non-headache migraine (acephalgic migraine). In some embodiments, the method comprises treatment of menstrual migraine. In a particular embodiment, the method of treating migraine comprises administering to an individual in need thereof an effective amount of N/OFQ or a pharmaceutical composition comprising N/OFQ intranasally.

In another aspect, provided herein is a method of treating a condition associated with migraine in an individual comprising administering (e.g., intranasally, buccally, sublingually or conjunctivally) an analgesic component (e.g., a non-opioid analgesic peptide, an NOP agonist or N/OFQ) or a pharmaceutical composition comprising an analgesic component (e.g., a non-opioid analgesic peptide, an NOP agonist or N/OFQ) via craniofacial mucosal administration. In some embodiments, the symptom is a prodromal stage symptom. In some embodiments, the symptom is a premonitory stage symptom. In some embodiments, the symptom is a pain stage symptom.

In some embodiments, the symptom is a late stage symptom. In some embodiments, the method comprises alleviating or preventing one or more of the symptoms associated with migraine, such as nausea, photophobia, and phonophobia. In a particular embodiment, the method comprises administering to an individual in need thereof an effective amount of N/OFQ or a pharmaceutical composition comprising N/OFQ nasally, wherein the administration reduces or prevents migraine-associated symptoms such as nausea, photophobia or phonophobia.

In certain aspects, the invention provides methods for treating pain by craniofacial mucosal administration (e.g., nasal, buccal, sublingual or conjunctival) of an analgesic component (e.g., a non-opioid analgesic peptide, a NOP agonist or N/OFQ). In some embodiments, the pain is somatic pain. In some embodiments, the pain is a superficial somatic pain. In some embodiments, the pain is a deep somatic pain. In some embodiments, the pain is a musculoskeletal pain. In some embodiments, the pain is a visceral pain. In some embodiments, the pain is a neuropathic pain. In some embodiments, the pain is a head pain or a craniofacial pain.

In some embodiments, the pain is a chronic pain, such as a chronic pain according to the present invention. In some embodiments, the pain is acute pain, one acute pain. In some embodiments, the pain is a combination of one or more of the pains described herein. In some embodiments, the pain is associated with a sharp tingling. In some embodiments, the pain is neuropathic pain resulting from surgery associated with a nerve injury.

Compound administration

In certain aspects of the invention, vasoconstrictors (vasoconstrictors) are used to reduce systemic absorption of analgesic components, such as non-opioid analgesic peptides, NOP agonists or N/OFQ. The vasoconstrictor may be included in a pharmaceutical composition to reduce systemic absorption of the analgesic component. In addition, the vasoconstrictor may be delivered to the mucosal or dermal surface separately from the pharmaceutical composition. Vasoconstrictors are compounds that constrict blood vessels and capillaries and reduce blood flow.

By inhibiting the passage of drugs into the blood, they can be used to increase the concentration of the drug at the intended site, thereby reducing the systemic absorption and distribution of the drug. By limiting the systemic distribution and concentration of the drug to the target tissues, i.e., cranial nerves and central nervous system, vasoconstrictors can be used to reduce the effective dose of the drug to achieve analgesia. Thus, the vasoconstrictor may be administered prior to or simultaneously with the analgesic peptide. Vasoconstrictors may include, but are not limited to, forline hydrochloride, tetrahydronaphthalene imidazoline (tetrahydrozoline), naphazoline hydrochloride nitric acid, oxymetazoline hydrochloride, tramazoline hydrochloride, ergotamine, dihydroergotamine, endothelin-1, endothelin, epinephrine, norepinephrine, and angiotensin.

In some embodiments, methods are provided that include administering a vasoconstrictor to the nasal cavity of the subject prior to administration of the analgesic component (e.g., a non-opioid analgesic peptide, a NOP agonist, or N/OFQ), wherein administration of the vasoconstrictor reduces the systemic distribution of the analgesic component. In some embodiments, the method may co-administer a vasoconstrictor and an analgesic component to the nasal cavity of the subject, wherein administration of the vasoconstrictor reduces the systemic distribution of the analgesic component.

In other examples, the method comprises administering a vasoconstrictor to the nasal cavity of the subject prior to administration of the analgesic composition, wherein administration of the vasoconstrictor reduces the systemic distribution of the analgesic composition, thereby reducing the effective dose of the analgesic composition to achieve analgesia.

In certain aspects, there is provided a method of treating pain in an individual, comprising: administering an effective amount of an analgesic component (e.g., a non-opioid analgesic peptide, NOP agonist or N/OFQ) or a pharmaceutical composition containing an analgesic component (e.g., a non-opioid analgesic peptide, NOP agonist or N/OFQ) via craniofacial mucosa, wherein the analgesic component is administered in combination with at least one additional active agent. In some embodiments, an analgesic component (e.g., a non-opioid analgesic peptide, a NOP agonist, or N/OFQ) is administered in combination with at least two additional active agents.

Additional active agents may include, but are not limited to, non-peptide opioids, such as morphine, methadone, fentanyl, butorphanol, codeine, opiates, oxycodone, loperimide, meperidine (ledum)) Diphenoxylate, propoxyphene (dar))4 methyl fentanyl, dihydrocodeine, morphine, diacetylmorphine, dihydrocodeine, hydrogenLevorphanol (levo)) Dextromethorphan, hydroxybenHeroin, fentanyl, benzathine, pentazocine, idazotine, ampicillin, buprenorphineSufentanil, carfenntanil, alfentanil and atypical opioids, tramadol, tapentadol; opioid and opioid peptides and analogs thereof, such as endorphins, enkephalins, dynorphins, picorphins, morphins, endomorphins and dalargin;

NMDA receptor antagonists such as ketamine, amantadine, dexromeorphine, memantine and MK801, such as local anesthetics and ergotamine sodium channel blockers, calcium channel blockers such as verapamil, nifedipine; adrenergic receptor antagonists, such as propranolol, metoprolol, and yohimine; GABAergic agonists such as aminobutyric acid, baclofen, cis-4-3-aminocrotonic acid (CACA), trans-4-aminocrotonic acid (TACA), CGP 27492 (3-aminopropophous acid) and progabide; glycine receptor agonists such as glycine and D-cycloserine; cholinergic agonists, such as neostigmine and physiostigmine; adrenergic receptor agonists, such as epinephrine, neosynephrine (neosynephrine), clonidine and dexmedetomidine, the anticonvulsants gabapentin and barbiturates; rho kinase inhibitors, such as fasudil, Y27632, H1152 and derivatives thereof; protein kinase C inhibitors such as chelerythrine, Go 6983, Go 6976, N-cardamom-serine-isoleucine-tyrosine-arginine-glycine-alanine-arginine-tryptophan-arginine-lysine-leucine, Rottlerin, KAI-9803 and KAI-1455, P38-MAP kinase inhibitors such as SCIO-469, AMG548 and derivatives thereof; ATP receptor blockers such as ligustrazine chelerythrine chloride A-317491 and derivatives thereof; endothelin receptor antagonists such as BQ123, BMS182874 and derivatives thereof; proinflammatory cytokines, chemokines, interleukins and tumor necrosis factor receptor blockers, such as anakinra, infliximab, etanercept and adalimumab, anti-inflammatory cytokines, such as interleukin 4, interleukin 10 and interleukin 13; such as tricyclic antidepressants, such as desiprimine and amitryptiline; serotonin antagonists such as fluoxetine, dolasetron and ondansetron; serotonin receptor agonists such as buspirone and ergometrine; NSAIDs and coxib drugs such as diclofenac, ibuprofen, ketorolac, salicylic acid, rofecoxib, celecoxib, parecoxib, valdecoxib and naproxen, acetaminophen; analgesic peptides such as calcitonin, octreotide, somatostatin, posterior pituitary, galanin, a lipotropin C-fragment and AC-rfwink-NH 2; toxins, e.g. botulinum toxin, variants and derivatives thereof, conotoxins, e.g. omega-dasheenSpiro toxin GV1A, omega-conotoxin MVIIA, shellfish toxin and tetrodotoxin; TPR channel agonists and antagonists, such as capsaicin, anti-capsaicin (capsazepine), resiniferatoxin, SB-705498, A-425619, AMG-517, SC0030 and derivatives thereof; cannabinoids such as HTC, CT-3, left-south-yeast dexanabinol, WIN-55, 212-2, AM 1241, dronabinol, nabilone, Cannabis Medicinal Extract (CME) and derivatives thereof; antagonists algesic peptide neurotransmitter receptors CGRP 1and CGRP2, including nonpeptidic antagonists such as BIBN4096 and its derivatives and peptide antagonists such as calcitonin, e.g., CGRP 8-37 and CGRP 28-3; NK1, an antagonist of algesin neurotransmitter receptors, including non-peptide antagonists such as SR140333, CP96346, L760735 RP 67580, WIN 51708; MK869, and derivatives and peptide antagonists thereof, e.g. N-acetyltryptophan, D-Pro9- [ Spiro-y-lactam]-Leu 10，Trp 11-Physalaemin(1-11)，Tyr-D-Phe-Phe-D-His-Leu-Met-NH₂(Sendaide) and spinode second analgesia antagonist peptide neurotransmitter receptor NK2, including non-peptide antagonists such as SR 48968 and derivatives and peptide antagonists thereof such as PhCO-Ala-Ala-D-Trp-Phe-D-Pro-Pro-Nle-NH₂(GR98400)[Tyr5，D-Trp6，8，9，Lys 10]-NKA (4-10) (MEN10376) and derivatives thereof; the parent pain peptide neurotransmitter receptor Y1-5 includes the nonpeptidic antagonists benextamine and peptide antagonists (ILE glue-PRO-DPR-tyrosine, arginine-leucine, arginine, tyrosine-NH 2)2, circulating (2, 4), (2, 4) diamine (1229U91 or GW1229), PYX 2D-tyrosine (27, 36) D-threonine (32)]Neuropeptide Y (27-36) (D-neuropeptide Y (27-36), 3- (5, 6, 7, 8-tetrahydro-9-isopropylcarbazol-3-YL) -1-methyl-1- (2-pyridin-4-YL-ethyl) urea hydrochloride (FMS586 and derivatives), neurotransmitter receptors VPAC2, VPAC1and PAC1 that are antagonists of nociceptive peptides, including peptide antagonists VI P (6-28), D-phenylalanine (2), K (15), R (16), L (27) of AC (1)]VIP (3-7)/GRF (8-27); the algenophilic receptor antagonist neurotransmitter receptors GAL1-3 and GalR1-3, SNAP including the non-peptide antagonists SNAP37889, 398299, galnon and derivatives thereof. Other active agents may include agonists or antagonists, vasopressin, Corticotropin Releasing Hormone (CRH), Growth Hormone Releasing Hormone (GHRH), Luteinizing Hormone Releasing Hormone (LHRH), growth hormoneSomatostatin growth hormone release inhibitory hormone (somatostatin), thyroid hormone releasing hormone (TRH), glial cell line-derived neurotrophic factor (GDNF), brain-derived neurotrophic factor (BDNF), Nerve Growth Factor (NGF), neurotrophic factor-3 (NT-3), pancreatic polypeptide, polypeptide tyrosine-tyrosine, glycogen-like peptide 1(GLP-1), histidine isoleucine peptide (philippine), pituitary adenylate cyclase activating peptide (PACAP's), brain natriuretic peptide, cholecystokinin (CCK), islet amyloid polypeptide (IAPP) or lake, Melanin Concentrating Hormone (MCH), melanocortins (adrenocorticotropic hormone, alpha-MSH and others), neuropeptide FF (F8Fa), neurotensin, parathyroid hormone-related protein, rat gene-related protein (AGRP), cocaine and amphetamine regulate transcription (order)/peptide, 5-hydroxytryptamine MODULINE, hypothalamic secretin/orexin, oxytocin peptide, ocistatin, prolactin releasing peptide, secretin, urocortin and its derivatives and analogs.

Pharmaceutical composition

Although it is possible to administer the analgesic component (e.g., a non-opioid analgesic peptide, NOP agonist or N/OFQ) alone, it is sometimes advantageous to administer it as part of a pharmaceutical composition. Thus, in certain aspects of the invention, an analgesic component (e.g., a non-opioid analgesic peptide, a NOP agonist or N/OFQ) is administered as a component of a pharmaceutical composition. Pharmaceutical compositions contain a therapeutically effective amount of an analgesic component (e.g., a non-opioid analgesic peptide, a NOP agonist or N/OFQ) in combination with one or more pharmaceutically acceptable carriers and optionally other ingredients.

A suitable carrier is one that does not cause intolerable side effects, but retains the pharmacological activity of the analgesic component (e.g., a non-opioid analgesic peptide, NOP agonist or N/OFQ) in vivo. The carrier may also reduce any adverse side effects of the analgesic component. A suitable carrier should be stable, i.e., generally non-reactive, with the other ingredients of the formulation. A suitable carrier has minimal unpleasant odor or aroma, but may include a pleasant aroma or a positive (pleasant) odor. A suitable carrier should not irritate the mucosa, epithelial cells, superficial nerves, causing health risks.

Suitable non-toxic pharmaceutically acceptable carriers will be apparent to those skilled in the art of pharmacy. See Remington: science and practice of pharmacy, 20 th edition, lipgkett williams & wilkins (2000). Typical pharmaceutically acceptable carriers include, but are not limited to, mannitol, dextran, urea, lactose, corn and potato starch, magnesium stearate, talc, vegetable oils, polyalkylene glycols, ethylcellulose, poly (vinyl), calcium carbonate, chitosan, ethyl oleic acid, isopropyl myristate, benzyl benzoate, sodium carbonate, gelatin, potassium carbonate, silicic acid, and other commonly acceptable carriers. Other carriers include, but are not limited to, lecithin, phospholipids, and sphingomyelins.

An appropriate carrier is selected depending on the exact nature of the particular dosage form desired, e.g., whether the drug is in the form of a liquid solution (e.g., as drops, for injection, as a spray or impregnated into nasal plugs, or other impregnated solids), a suspension, ointment, film, or gel. If necessary, it can be prepared into sustained-release gel, film, scalp patch, etc. The particular dosage form will also depend on the route of administration. The medicament may be administered to the nasal cavity as a powder, granule, solution, cream, spray, gel, film, ointment, infusion, drop or sustained release composition. The oral administration of the composition may be in the form of tablets or lozenges. For sublingual administration, the composition may be in the form of a bioadhesive, spray, powder, paint or a cotton swab for lingual or sublingual administration. The composition can be administered as an ointment, solution or drops to the conjunctiva or other mucosal tissue surrounding the eye. For dermal administration, the composition may be applied as a topical ointment, topical gel, lotion, cream, solution, spray, paint, film, foil, cosmetic, plaster or bioadhesive.

Liquid carriers for making solutions include, but are not limited to, water, physiological saline, aqueous dextrose, and particularly glycol (isotonic) solutions. The carrier can also be selected from a variety of oils including those of petroleum, animal, vegetable or synthetic origin (e.g., peanut oil, soybean oil, mineral oil, sesame oil, and the like). Suitable pharmaceutical excipients include, but are not limited to, starch, talc, cellulose, glucose, lactose, sucrose, gelatin, malt, rice, flour, chalk, silica gel, magnesium stearate, sodium, stearic acid, glycerol monostearate, sodium chloride, skim milk, glycerol, propylene glycol, water, ethanol, and the like. The pharmaceutical composition may be subjected to conventional pharmaceutical processes such as sterilization, and may contain conventional pharmaceutical additives such as preservatives, stabilizers, reducing agents, antioxidants, chelating agents, wetting agents, emulsifiers, dispersing agents, gelling agents, salts for adjusting osmotic pressure, buffers and the like. If the carrier is a liquid, the carrier is hypotonic or isotonic with body fluids, at a pH in the range of 4.5-8.5. If the carrier is in powder form, the carrier is preferably within an acceptable, non-toxic pH range. The use of additives in peptide and/or protein based compositions, particularly pharmaceutical compositions, is well known in the art.

The carriers and additives listed in the present invention are not all inclusive and those skilled in the art can select carriers and excipients from the GRAS (generally recognized as safe) pharmaceutical acceptable chemical list and those currently acceptable and injectable formulations. (see also Wang et al, (1980) J.Parent.drug Assn., 34: 452-462; Wang et al, (1988) J.Parent.Sci.and Tech., 42: S4-S26.)

Other dosage forms of the composition include suspensions of microparticles, such as emulsions, liposomes, or in a sustained release form, which prolong the presence of the individual pharmaceutically active ingredients. Powdered or granular forms of the pharmaceutical compositions may incorporate solvents, diluents, dispersing agents or surfactants. In addition, additional components include biological adhesives to keep the drug immobilized at the site of administration, e.g., sprays, paints, or swabs applied to the mucosa or epithelial cells. A bioadhesive may be a hydrophilic polymer, natural or synthetic, wherein the hydrophilic formulation, which may be water soluble or water swellable, is compatible with the pharmaceutical ingredients.

Such adhesives function to adhere the drug to the mucosal tissue of the oral or nasal cavity. Such binders may include, but are not limited to, hydroxyethyl cellulose, hydroxypropyl methyl cellulose, ethyl cellulose, carboxymethyl cellulose, dextran, oxhide gum, polyvinyl pyrrolidone, pectin, starch, gelatin, casein, acrylic acid polymers, acrylate polymers, acrylic acid copolymers, ethylene polymers, ethylene copolymers, vinyl alcohol polymers, alkoxy polymers, polyethylene oxide polymers, polyethers, and combinations thereof. Can also be made into lyophilized powder, and can be converted into solution, suspension, or emulsion before administration. The pharmaceutical compositions are preferably sterile membrane filtered and stored in unit-dose or multi-dose containers, or sealed vials or ampoules.

The pharmaceutical compositions may be formulated in sustained release form to prolong the presence of the active ingredient in the treated individual. Sustained release formulations are well known in the art and are described in the pharmaceutical science of Remington (see above). In general, the drug may also be embedded in a semipermeable solid hydrophobic polymer matrix. The matrix may be formed into a film or microcapsules. The matrix may include, but is not limited to, polyester, L-glutamic acid and gamma ethyl-L-glutamic acid, polylactic acid polyglycolic acid (polylactate polyglycolite), hydrogel, non-degradable ethylene-vinyl acetate, degradable polymers of lactic acid-glycolic acid copolymers, hyaluronic acid gel, alginic acid suspension.

Suitable microcapsules include amino acids (hydroxymethyicellulose) or gelatin and polymethylmethacrylate. Microemulsion or colloidal drug delivery systems, such as liposomes and albumin microspheres, may also be used. Some slow release components may be held in place using a bioadhesive.

To further enhance delivery of pharmaceutical compositions containing an analgesic component (e.g., a non-opioid analgesic peptide, NOP agonist or N/OFQ), enzyme inhibitors, particularly protease inhibitors, may be included in the dosage form. Protease inhibitors may include, but are not limited to, antipain, Oximenine (orphaminine) Aand B, benzamidine HCl, AEBSF, CA-074, Calpain inhibitors I and II, calpeptin, pepstatin, actinonin, aminopeptidase inhibitors, peptidase inhibitors, boroleucine (boroleucin), captopril, chloroHO leucine-alanine-glycine-NH 2, DAPT, aprotinins A and B, ebisinolide A and B, formoxyshipine, leupeptin, pepstatin A, amiloride, aprotinin, puromycin, BBI, soybean trypsin inhibitor, phenylmethylsulfonyl fluoride, E-64, chymotrypsin inhibitor, 1, 10-phenanthroline, EDTA, and EGTA.

To enhance delivery or passage across mucosal surfaces and/or absorption of pharmaceutical compositions containing analgesic components (e.g., non-opioid analgesic peptides, NOP agonists or N/OFQ), absorption-promoting components may be included in the dosage form. The absorption-promoting component can enhance the release or dissolution of the drug (e.g., from the delivery vehicle), diffusivity, permeability and timing, absorption, residence time, stability, effective half-life, peak or sustained concentration levels, clearance, and other desirable mucosal delivery characteristics (e.g., measurement of delivery site).

Thus, enhanced mucosal delivery may be achieved by various mechanisms, for example, by increasing the diffusion, transport, persistence or stability of an analgesic component (e.g., a non-opioid analgesic peptide, NOP agonist or N/OFQ), increasing membrane fluidity, modulating the availability or activity of calcium and other ions, modulating intracellular and extracellular osmotic pressure, solubilizing mucosal components (e.g., blood lipids), altering protein and non-protein sulfhydryl levels in mucosal tissue, increasing water flux across mucosal surfaces, modulating the physiology of epithelial tissue interfaces, reducing the viscosity of mucus overlying mucosal epithelium, reducing ciliary clearance, and other mechanisms.

Compounds that enhance mucosal absorption include, but are not limited to, surfactants, bile salts, dihydrofusidates, bioadhesives, phospholipid additives, mixed microparticles, liposomes, or carriers, alcohols, enamines, cationic polymers, NO donor compounds, long chain amphiphilic molecules, low hydrophobic transdermal absorption enhancers, sodium or salicylic acid derivatives, glyceryl acetoacetate, cyclodextrin or β -cyclodextrin derivatives, medium chain fatty acids, chelating agents, amino acids or salt derivatives thereof. N-acetylamino acid or its salt derivative, expectorant, enzyme selected specifically for membrane component, and inhibitor of fatty acid synthesis and cholesterol synthesis.

These agents and compounds may be administered in conjunction with or combined with analgesic ingredients (e.g., peptides other than opioid analgesics, NOP agonists or/OFQ). Accordingly, aspects of the invention include methods of administering an analgesic component (e.g., a peptide other than an opioid analgesic, an NOP agonist or/OFQ) as a pharmaceutical composition comprising a protease inhibitor, an absorption enhancer, a vasoconstrictor, or combinations thereof.

The pharmaceutical composition can be administered to the nasal cavity, oral cavity, conjunctiva or other tissues around the eyes or skin mucosa. The pharmaceutical composition may be administered by the nasal route. The pharmaceutical composition may be administered by the buccal or sublingual route. The pharmaceutical composition can be administered by the transdermal route. The pharmaceutical composition may be administered by taking more than one route. The pharmaceutical composition may include at least one protease inhibitor, at least one absorption enhancer, at least one vasoconstrictor, or a combination thereof. The pharmaceutical composition may be administered simultaneously with the vasoconstrictor or may be administered after the administration of the vasoconstrictor.

Dosage form

An analgesic component (such as a peptide other than an opioid analgesic, a NOP agonist or/OFQ) is administered in a sufficient dose to effectively treat an individual suffering from pain. In certain aspects, analgesic ingredients (e.g., peptides other than opioid analgesics, NOP agonists or/OFQ) may be administered via the craniofacial mucosal route (e.g., the nasal route) at doses that result in systemic analgesic effects with minimal central nervous system or systemic side effects. A therapeutically effective dose of an analgesic component (e.g., a peptide other than an opioid analgesic, a NOP agonist or/OFQ) can be determined by one of skill in the art, such as a pharmacist, empirically and with the type and severity of the pain, the route of administration, and according to the patient's size, weight, age, and general health.

The amount of analgesic ingredient (e.g., a peptide other than an opioid analgesic, NOP agonist or/OFQ) administered as a unit dose depends on the type of pharmaceutical composition being administered, e.g., a solution, suspension, gel, film, emulsion, powder, or sustained release formulation. In some instances, the effective dose is lower than that required for oral, intravenous, intramuscular or subcutaneous injection, as intramuscular injection or transdermal administration may result in a greater concentration of the analgesic component in the craniofacial or cephalic region. The amount of formulation required to deliver the required dose will also depend on the concentration of the analgesic component in the pharmaceutical composition. Such a determination is within the skill of the art.

The therapeutic dosage of the pain-stopping component (e.g., a peptide other than an opioid analgesic, a NOP agonist or/OFQ) used in the methods of the present invention depends on a number of factors, such as the modification of the chemical composition and/or the analgesic component, the bioavailability of the route of administration, the therapeutic efficacy, the frequency of the combined administration of the individual doses of the formulated formulation, and whether the analgesic component is combined with other active agents.

In particular, the dosage of analgesic component (e.g., a peptide other than an opioid analgesic, NOP agonist or/OFQ) is required to minimize pain in the patient being treated. Pharmaceutical data can be collected by a number of operators in the field from animal models and clinical trials, and normal human volunteers or patient experiences.

Experimental models for testing the analgesic effect of drugs are well known in the art. Animal models, including tests, including, but not limited to, acetate writhing, benzoquinone writhing, tail flick, withdrawal of paw and acetate, benzoquinone induction, formalin, capsaicin activation, or withdrawal when heat such as hot plate or laser activates the cheek/ear/face pain receptors. In particular, models for craniofacial or cephalic pain testing, such as facial applications of capsaicin, facial applications of formalin, or heat transfer to the cheek, ear, or face, are possible.

In some cases, other models may be used to develop an analgesic effect test, such as the models described in the present invention. The model can be used to determine the optimal dosage range of analgesic effect of the analgesic component with minimal central nervous system and/or systemic side effects. In addition, the model may be used to administer the analgesic composition via a specific delivery route, such as nasal drops, or to test the analgesic effect on the cheek, ear or face or hind paw. In some cases, a model may be used to test the analgesic activity of the analgesic component after administration of the pharmaceutical composition, and to manage the pharmaceutical composition, wherein the analgesic activity of the analgesic agent after withdrawal latency in the cheek, ear or face may determine regional analgesia, while the withdrawal latency may determine the systemic analgesic effect.

As noted above, the effective amount of analgesic component (e.g., a non-opioid analgesic peptide, a NOP agonist or N/OFQ) will depend on the form and composition of the method used. An effective dose of the N/OFQ peptide is administered via craniofacial mucosa at a lower dose than that used by other routes of administration (e.g., oral, intravenous, intramuscular, or subcutaneous injection), which may not be effective at all, or may require a higher dose. For example, a dosage for an N/OFQ peptide can include, but is not limited to, from about 0.2mg to about 5000mg, from about 0.2mg to about 2000 mg, from about 0.2mg to about 1000 mg, from about 0.2mg to about 200 mg, from about 0.2mg to about 100mg, from about 0.5 mg to about 100mg, or from about 0.5 mg to about 50 mg, from about 0.2mg to about 500 mg, or from about 0.5 mg to about 25mg, or from about 0.5 mg to about 10mg, or from about 0.5 mg to about 5mg, or from about 0.5 mg to 1mg, or from about 1mg to about 100mg, or from about 1mg to about 50 mg, or from about 1mg to about 25mg, or from about 1mg to about 10mg, or from about 1mg to about 5mg, or from about 5mg to about 200 mg, or from about 5mg to about 100mg, about 5mg to about 50 mg, about 5mg to 25mg, about 5mg to 10 mg. The amount may be administered in a single dose or in multiple doses, for example, the dose may be administered twice, three times, four times daily, up to ten times, depending on the type and severity of the headache, and the sensitivity of the patient. The amounts may be administered in a slow release formulation such that the N/OFQ peptide is administered less frequently, e.g., 6 times per week, 5 times per week, four times per week, three times per week, twice per week, or once per week.

Accordingly, certain aspects of the invention include methods of treating pain by administering to a subject via nasal administration an effective amount of an N/OFQ peptide. The dosage range for the N/OFQ peptide to be administered can be determined by methods well known in the art for animal models and/or human clinical trial studies.

For example, a unit dose may range from 0.2mg to 5000mg, from about 0.2mg to about 2000 mg, from about 0.2mg to about 1000 mg, from about 0.2mg to about 200 mg, or from about 0.2mg to about 100mg, or from about 0.5 mg to about 50 mg, from about 0.2mg to 500 mg, or from about 0.5 mg to 25mg, or from about 0.5 mg to about 10mg, or from about 0.5 mg to about 5mg, or from about 0.5 mg to 1mg, or from about 1mg to about 100mg, or from about 1mg to about 50 mg, or from about 1mg to about 25mg, or from about 1mg to about 10mg, or from about 1mg to about 5mg, or from about 5mg to about 200 mg, or from about 5mg to about 100mg, from about 5mg to about 50 mg, about 5mg to about 25mg, about 5mg to about 10 mg.

In certain aspects of the invention, a composition comprising an analgesic component (e.g., a non-opioid analgesic peptide, a NOP agonist, or N/OFQ) further comprises an additional active agent, wherein the analgesic component and the additional active agent are administered as a mixture, separately, simultaneously or separately, in any order. In some embodiments, the N/OFQ peptide-containing composition is administered in combination with at least one additional active agent. In other embodiments, the composition comprising the N/OFQ peptide is administered in combination with at least 2 additional active agents. In other embodiments, the composition comprising the N/OFQ peptide is administered in combination with diclofenac, oxytocin, or an oxytocin receptor antagonist.

To determine the therapeutic efficacy of an analgesic component (e.g., a non-opioid analgesic peptide, NOP agonist or N/OFQ) or a pharmaceutical composition containing an analgesic component (e.g., a non-opioid analgesic peptide, NOP agonist or N/OFQ), a "visual analog score" (VAS) can be used to assess the reduction or alleviation of pain following administration of the analgesic component. VAS is a 10 cm turn point, either horizontal or perpendicular to the end, such as "painless" and "extremely painful". A subject or patient needs to make an indication of the level of pain at the line.

This designation will be converted to a distance in centimeters (cm) or millimeters (mm) from "painless", and pain will be scored, which may be from 0-10 cm or 0-100 mm. The VASX is similar to an 11-point value pain rating scale, where 0 equals "no pain" and 10 equals "most pain imaginable,", or vice versa. With VAS, an analgesic component, such as a non-opioid analgesic peptide, NOP agonist or N/OFQ, is considered to have a clinically relevant analgesic effect when there is a change of about 30% or more, for example, from 9 to 7 or 5 to 3.5.

Drug delivery system

An analgesic component (e.g., a non-opioid analgesic peptide, NOP agonist or N/OFQ) or a pharmaceutical composition containing an analgesic component (e.g., a non-opioid analgesic peptide, NOP agonist or N/OFQ) may be delivered to the oral or sublingual surface in various dosage forms or formulations including, but not limited to, fast-melt tablets, liquid-filled capsules, liquid sprays or lozenges. Alternatively, the pharmaceutical composition may be delivered to the mucosa of the oral cavity by direct placement in the mouth, by means of, for example, a gel, film, ointment, dropper, or bioadhesive or patch.

In certain aspects of the invention, the method of administering a pharmaceutical composition to a subject to the buccal or sublingual mucosa may be administered by an administration device. Drug delivery devices may include, but are not limited to, unit dose containers, spray pumps, droppers, squeeze bottles, vacuum and preservative-free nebulizers, nebulizers and pressurized inhalers. The administration device can meter the accurate effective dose to the oral cavity. In certain aspects, an accurate effective dose is contained in a capsule, tablet, troche, or bioadhesive patch placed directly in the mouth.

An analgesic component (e.g., a non-opioid analgesic peptide, NOP agonist or N/OFQ) or a pharmaceutical composition containing an analgesic component (e.g., a non-opioid analgesic peptide, NOP agonist or N/OFQ) can be applied to the conjunctiva or other mucosal tissue surrounding the eye in various dosage forms, such as a drop of liquid, gel, film, ointment, or bioadhesive patch or bandage. Thus, in certain aspects, the methods of the invention comprise administering a pharmaceutical composition to a subject, wherein the administration is directly to the conjunctiva versus other mucosal tissues surrounding the eye. In certain aspects, an accurate effective amount is contained in drops, gels, films, ointments, or bioadhesive patches, administered directly to the mucosal tissue surrounding the eye.

An analgesic component (e.g., a non-opioid analgesic peptide, NOP agonist or N/OFQ) or a pharmaceutical composition containing an analgesic component (e.g., a non-opioid analgesic peptide, NOP agonist or N/OFQ) can be administered to the skin or scalp in a number of different dosage forms, such as a liquid, spray, gel, film, ointment or bioadhesive patch or bandage. Thus, in certain aspects, the methods of the invention comprise administering a pharmaceutical composition to an individual, wherein administration is directly to the skin, e.g., the anterior scalp. In certain aspects, an accurate effective amount is contained in drops, gels, films, ointments, or bioadhesive transdermal patches, which are administered directly to the skin. In certain aspects, the pharmaceutical composition may be injected intradermally into the skin. In other aspects, the pharmaceutical composition can be injected subcutaneously into the skin.

An analgesic component (e.g., a non-opioid analgesic peptide, NOP agonist or N/OFQ) or a pharmaceutical composition containing an analgesic component (e.g., a non-opioid analgesic peptide, NOP agonist or N/OFQ) can be dispensed intranasally as a powder or liquid nasal spray, suspension, nasal drop, gel, film or ointment, by tube or catheter, syringe, packtail, gauze (a small unit absorbent pad), nasal tampon, or by infusion into the submucosa.

Nasal administration can be carried out using devices including, but not limited to, unit dose containers, spray pumps, droppers, squeeze bottles, vacuum and preservative-free nebulizers, nebulizers (devices used to modify liquid drugs into aerosol particulate form), metered dose inhalers, and pressurized metered dose inhalers. It is important that the drug delivery device protect the drug from contamination and chemical degradation. The device should also avoid leaching or absorption and provide a suitable storage environment. Each drug requires an evaluation to determine which nasal delivery devices are most suitable. Nasal delivery systems are well known in the art and are also commercially available.

An analgesic component (e.g., a non-opioid analgesic peptide, NOP agonist or N/OFQ) or a pharmaceutical composition containing an analgesic component (e.g., a non-opioid analgesic peptide, NOP agonist or N/OFQ) is conveniently administered in the form of an aerosol spray using a pressurized pack or nebulizer and a suitable propellant, including, but not limited to, dichlorodifluoromethane, trichlorofluoromethane, dichlorotetrafluoroethane, hydrocarbons, nitrogen or carbon dioxide, compressed air. Aerosol systems require that the propellant be inert to the pharmaceutical ingredients. In the case of a pressurized aerosol, the dosage unit can be controlled to the exact unit by setting a valve.

An analgesic component (e.g., a non-opioid analgesic peptide, NOP agonist or N/OFQ) or a pharmaceutical composition containing an analgesic component (e.g., a non-opioid analgesic peptide, NOP agonist or N/OFQ) is administered to the nasal cavity, e.g., as a powder, or pressurized in the form of microspheres administered via a nasal insufflator device (insufflation of air, powder or vapor in the cavity) or aerosol canister. The insufflator may produce finely divided dry powders or microspheres.

The insufflator is required to ensure that a sufficient amount of the pharmaceutical composition is provided for administration. The dry powder or microspheres should be administered in a dry, air-free or otherwise. The dry powder or microspheres can be directly administered by an insufflator which can spray the dry powder or microspheres on a medicine bottle or a container. Alternatively, the dry powder or microspheres may be filled into a capsule, such as a gelatin capsule, or other single dose device suitable for nasal administration. The insufflator may be used to administer a powdered composition to the nasal cavity by opening a capsule or other means of providing a throat-like device with a needle.

Nasal delivery devices may be constructed or modified to deliver an analgesic component (e.g., a non-opioid analgesic peptide, NOP agonist or N/OFQ) or a pharmaceutical composition containing an analgesic component (e.g., a non-opioid analgesic peptide, NOP agonist or N/OFQ) wherein the analgesic component or composition is delivered primarily to less than two-thirds of the nasal cavity. For example, the angle of emanation from a delivery device such as a nebulizer or insufflator may be such that the pharmaceutical composition is mechanically administered to less than two thirds of the nasal cavity and away from the upper nasal end region.

Alternatively, the analgesic component (e.g., a non-opioid analgesic peptide, NOP agonist or N/OFQ) or a pharmaceutical composition containing the analgesic component (e.g., a non-opioid analgesic peptide, NOP agonist or N/OFQ) may be placed directly into the nasal cavity for delivery to less than two-thirds of the nasal cavity, for example, in a gel, ointment, nasal tampon, dropper, or bioadhesive strip. Alternatively, an analgesic component (e.g., a non-opioid analgesic peptide, NOP agonist or N/OFQ) or a pharmaceutical composition containing an analgesic component (e.g., a non-opioid analgesic peptide, NOP agonist or N/OFQ) may be administered to the upper nasal cavity region, or both the upper and lower nasal cavities may be administered together.

Thus, in certain aspects of the invention, the method comprises administering to a subject an analgesic component (e.g., a non-opioid analgesic peptide, NOP agonist or N/OFQ) or a pharmaceutical composition comprising an analgesic component (e.g., a non-opioid analgesic peptide, NOP agonist or N/OFQ), wherein administration to the nasal cavity is via a nasal delivery device.

Nasal delivery devices may include, but are not limited to, unit dose containers, spray pumps, droppers, squeeze bottles, vacuum and preservative-free nebulizers, aerosol and pressurized inhalers, insufflators, or bi-directional devices. Nasal delivery devices can meter the amount of an effective dose to be administered nasally (as described below). Nasal delivery devices may be single dose units, or multiple dose units. Nasal delivery devices may be delivered as a single unit or multiple units. In some embodiments, the nasal delivery device may be constructed such that the angle of emanation of the pharmaceutical composition is mechanically directed to less than two thirds of the nasal cavity, thereby minimizing delivery to the olfactory site. In some embodiments, the nasal delivery device may be constructed such that the angle of emanation of the pharmaceutical composition is mechanically directed to the upper nasal cavity.

In some embodiments, the nasal delivery device may be constructed such that the angle of emanation of the pharmaceutical composition is mechanically directed to the lower and upper nasal cavity regions. In some embodiments, the nasal delivery device may be activated only upon exhalation, thereby limiting the induction of inhalation and potential undesired distribution of the pharmaceutical composition. In some embodiments, the pharmaceutical composition is a gel, film, cream, ointment, nasal tampon, or bioadhesive strip, wherein the pharmaceutical composition is placed in the nasal cavity under two-thirds, in the upper nasal cavity, or both.

In some embodiments, the method comprises nasally administering an analgesic composition (e.g., a non-opioid analgesic peptide, an NOP stimulant or N/OFQ) or a pharmaceutical composition comprising an analgesic composition (e.g., a non-opioid analgesic peptide, an NOP stimulant or N/OFQ), wherein the administration is via a nasal delivery device that mechanically directs the drug to less than two-thirds of the nasal cavity, to the upper nasal cavity, or both, after administration of the vasoconstrictor.

In some embodiments, the method comprises nasally administering an analgesic agent (e.g., a non-opioid analgesic peptide, an NOP stimulant, or N/OFQ) or a pharmaceutical composition comprising an analgesic agent (e.g., a non-opioid analgesic peptide, an NOP stimulant, or N/OFQ), in conjunction with a vasoconstrictor, wherein the administration is via a nasal delivery device that mechanically directs the agent to the nasal cavity at an angle that is less than two times the angle of divergence of the agent, to the upper nasal cavity, or both.

Reagent kit

The invention provides kits for carrying out the methods. The kit is for use in the treatment and/or prevention of pain. In some embodiments, the kit includes an analgesic component (e.g., a non-opioid analgesic peptide, an NOP agonist, or N/OFQ) in suitable packaging. In some embodiments, the kit comprises a suitable package of N/OFQ peptide. In some embodiments, the kit further comprises at least one additional analgesic component. The kit may further comprise a vasoconstrictor, at least one protease inhibitor and/or at least one absorption enhancer. Some kits may further include drug delivery devices, including, but not limited to, nasal drug delivery devices. Other kits may further include instructions for providing information to the user and/or health care provider to perform the methods.

Kits comprise a single component, e.g., an analgesic component, e.g., a non-opioid analgesic peptide, NOP agonist, or N/OFQ, typically sealed within a container (e.g., a vial, ampoule, or other suitable storage container). Likewise, a kit comprising multiple components also comprises the components in a single container (either separately or mixed).

With regard to instructions for using the kits for carrying out the invention, it is generally described how to use the contents of the kit to carry out the methods described herein. The instructions provided by the kits of the invention are typically written on a label or package insert (e.g., paper in the kit), and machine-readable instructions (e.g., instructions on a magnetic or optical storage disk) are also acceptable.

Examples

The present invention will be further understood from the following examples, which are provided in connection with the accompanying drawings, and the invention is not intended to be so limited.

Example 1

The method of testing the activity of analgesic ingredients in rat models is to induce changes in treatment by the withdrawal latency (time) of the thermal stimulus response to the skin, typically using the ear, face or, hind limb. Thus, the application of continuous or discontinuous (non-laser) radiant heat to the ear, face or hind limb will cause rapid retraction behaviour. Withdrawal latencies have been shown to be sensitive to analgesic treatments, such as analgesics increasing withdrawal latencies.

Administration of transmucosal or transdermal analgesics to the trigeminal nerve to reduce trigeminal-related pain can be used to measure regional and/or systemic analgesic effects of the trigeminal nerve. The mouse's ear medulla external part is dominated by a branch of the mandibular nerve, which is itself a branch of the trigeminal nerve. Thus, an increase in withdrawal latency time following treatment indicates regional analgesia. Similarly, changes in hind limb withdrawal latency time indicate a systemic analgesic effect.

Rats were housed in an 12/12 hour light/dark environment and were provided with food and water ad libitum. The number of animals was minimized to reduce discomfort. Rats were gently anesthetized with urethane and placed on a minimally restrictive heating pad to maintain their own body temperature at 37 □ C. The laser beam is directed through the fiber optic cable to the outside of the two ear medulla. The response to laser irradiation is characterized by the withdrawal or withdrawal of the stimulated ear after 1-3 seconds of laser stimulation. Laser stimulation is terminated rapidly after the stimulated ear response, or 30 seconds of latency past a maximum response (nodal point) to prevent tissue damage.

In a baseline test of withdrawal latency of ear response, 3 pulses were applied to each ear trigeminal innervated segment. After at least 2 minutes of stimulation on the same ear with each pulse, the stimulation site was changed. In baseline testing of hindlimb response withdrawal latency, 3 pulses were applied to the hindlimb. After at least 2 minutes of stimulation on the same hindlimb per pulse, the stimulation site was changed. The test will be recorded for off-line reaction analysis. Off-line analysis was performed by researchers who determined the laser stimulus response withdrawal latency and knowledge of treatment group status.

After measuring the latency baseline, the analgesic component was administered intranasally. This involved 5 nasal drops of 10 microliter total volume of 50 μ L of drug over 20 minutes. The effect of withdrawal latency (e.g., 5mg/kg nitrogen N/OFQ) for different doses of drug was tested. To assess local analgesic effects, the latency response of the ear was tested at various time points after administration. To assess systemic effects, latency responses in the hind paw were tested at various time points after dosing.

Example 2: analgesic effect of intranasal analgesic heat injury experiment

A thermally nociceptive rodent model was used to determine the dose of nasal drops of the nociceptin in relation to 1) withdrawal latency, 2) persistence, 3) providing evidence to support or oppose the hypothesis that the analgesic effect is centrally mediated by the nociceptin.

Apparatus and materials

Nine male SD rats per group (200- & ltSUB & gt 300G) were treated with pain-causing substances dissolved in saline (i.n. used at concentrations of 0.5 mg/ml, 5.0 mg/ml, 50 mg/ml), i.v. four pain-causing substances (50 mg/ml), morphine (1.25 mg/ml), saline, polyurethane, A-delta/C pain detector, timer.

Reference to the literature

Yeomans，D.C.Pirec，V.，and Proudfit，H.K.“Nociceptive responsesto high or low rates of noxious cutaneous heating are mediated bydifferent nociceptors in the rat：behavioral evidence”Pain(1996)68：133-140.

Sherman S.E，Loomis C.W.“Morphine Insensitive Allodynia Is ProducedBy Intrathecal Strychnine in the Lightly Anesthetized Rat”Pain(1994)56：17-29.

Thorne R.G.，Pronk G.I，Padmanabhan V，Frey W.H.2nd.“Delivery ofInsulin-Like Growth Factor-I to the Rat Brain and Spinal Cord alongOlfactory and Trigeminal Pathways following IntranasalAdministration”Neuroscience(2004)127：481-96.

Yeomans，D.C.Pirec，V.，and Proudfit，H.K.“Nociceptive responsesto high or low rates of noxious cutaneous heating are mediated bydifferent nociceptors in the rat：behavioral evidence”Pain(1996)68：133-140.

Sherman S.E，Loomis C.W.“Morphine Insensitive Allodynia Is ProducedBy Intrathecal Strychnine in the Lightly Anesthetized Rat”Pain(1994)56：17-29.

Thorne R.G.，Pronk G.I，Padmanabhan V，Frey W.H.2nd.“Delivery ofInsulin-Like Growth Factor-I to the Rat Brain and Spinal Cord alongOlfactory and Trigeminal Pathways following IntranasalAdministration”Neuroscience(2004)127：481-96.

Contents of the experiment

The salts are lyso-algesin, morphine and physiological saline.

Test protocol/flow diagram

(A) 6 rats per test group were lightly anaesthetised with polyurethane (1.0g/kg, i.p., with minimal active titration in the absence of irritation) (Sherman and Loomis et al, 1996). The left and right cheeks and the right hind limb were blackened with ink to distribute the heat evenly.

(B) Different levels of thermal pain intensity A-delta were compared to C-fiber tests (Yeomans et al, 1996) on the left, right cheek and right hind limb of 200-300G male SD rats.

In the C-fiber experiment, the heat intensity was adjusted by varying the power supply voltage of the focused projector lamp until a retraction latency of between 9 and 13 seconds was observed. The intensity (in volts) applied to each animal was recorded. To reduce potential tissue damage, a 20 second latency period served as a node to stop stimulation. Rats that did not respond to a 55 volt supply voltage were not included in this study.

For the a-delta fiber experiment, the heat intensity was adjusted until the observed withdrawal latency occurred in 2 to 3 seconds. The intensity (volts) applied to each animal was recorded. To reduce potential tissue damage, the 6 second heat generation time served as a node to stop stimulation. Rats that did not respond to a supply voltage of 85 volts were not included in the study.

(C) Rats were treated with one of the following:

48 μ L of saline or nociceptin (0.2, 1.0 and 5.0mg/kg) was applied to the nose (Frey Method, see below; Thorne et al, 2004).

50 microliters of intravenous saline or analgesic (5.0mg/kg)

50 μ L of i.m. saline or morphine (0.25mg/kg)

The experimental animals were treated by eye-covering.

(D) The withdrawal latency for each hind limb to the radiant heat stimulus was measured after the withdrawal latency for the left, right cheek radiant heat stimulus response. Each animal was tested for its specific radiation amplitude (volts) established in the baseline experiment. Detection was performed immediately after dosing and at 15, 30, 45, 60, 90, 120, 150 and 180 minutes.

(E) Rats were euthanized by CO2 inhalation.

A Frey method:

the rat is placed on the back, the pad is inserted into the supporting surface from the back to the neck until the back of the head, and the whole process of the upper surface of the neck and the supporting surface is kept horizontal. (6) microliter of liquid containing the peptide, every 2 minutes into the nasal cavity with a small pipette (every four minutes of dose in the same nostril), with alternate nasal drops on either side. Each nostril received 4 doses of 6 μ l peptide-containing liquid,

two problems are likely if nasal administration of anesthetized rats is more rapid than described above: first, mice may have respiratory distress because the drops do not have time to be completely absorbed and eventually absorbed through the nasopharynx into the lungs; second, less drug delivery results because the previously delivered liquid already covers the nasal mucosa.

In this experiment, rats were anesthetized with urethane (urethane).

Figures 1and 3 show the analgesic effect of nasal dosing of rat nociceptin compared to the a-delta pain threshold test for intravenous nociceptin, morphine doses under general anesthesia or nasal drops of physiological saline. In fig. 1, the Y-axis shows the latency of facial (cheek) withdrawal in rats in response to thermal stimuli, selectively activating the components of the pain system of the a-delta fiber receptor that are subject to intense pain. The Y-axis in figure 3 shows the paw withdrawal latency of the rat hind limb in response to thermal stimuli, selectively activating the a-delta fiber receptor. The X-axis indicates the time the analgesic effect is maintained after administration.

Figures 2 and 4 show the dose-dependent analgesic effect of nasal administration of rat nociceptin compared to the a-delta pain threshold test for intravenous injection of buprenorphine, systemic anesthetic morphine doses or normal saline nasal drops. In figure 2 the Y-axis shows the cheek retraction latency of rats in response to thermal stimulation, selectively activating the C-fiber receptor-pain system component for burning pain. The Y-axis in fig. 4 shows the paw withdrawal latency in response to thermal stimuli, with the paw selectively activating the pain system component of the C-fiber receptors for burning pain at one rate. The X-axis indicates the time the analgesic effect is maintained after administration.

The rat face and hind limb A-delta and the C-fiber fibroreceptor response to heat activation after nasal nociceptin (N/OFQ) is well-known as dose-dependent. Nasal drops of physiological saline and intravenous pain-producing hormone (/ OFQ) did not show a response of the face and hind limb A-delta and the C-fiber receptors to heat-activated behavior.

Example 3:effect of antagonists of the painkiller receptor on the reduction of thermal pain sensation in the nasal cavity caused by the painkiller

This experiment was to verify the binding site for the analgesic effect of the analgesic in rodent models of thermal pain sensation.

I. Apparatus and materials

Nine male SD rats per group (200-300G), with oxytocin dissolved in saline (50 mg/ml), nociceptin receptor antagonist (SB-612111{ (-) -cis-1-methyl-7- [ [4- (2, 6-dichlorophenyl) piperidine-1-yl ] methyl ] -6, 7, 8, 9-tetrahydro-5H-benzocyclohepten-5-ol }, 100mg/ml), saline, polyurethane, A-delta/C algesin, chronograph.

Reference II

The same references as in example 2, and

Chiou LC，Liao YY，Fan PC，Kuo PH，Wang CH，Riemer C，Prinssen EP.“Nociceptin/orphanin FQ peptide receptors：pharmacology and clinicalimplications”Curr.Drug Targets(2007)8(1)：117-35.

Spagnolo B，Carra G，Fantin M，Fischetti C，Hebbes C，McDonald J，BarnesTA，Rizzi A，Trapella C，Fanton G，Morari M，Lambert DG，Regoli D，CaloG.“Pharmacological Characterization of the Nociceptin/Orphanin FQReceptor Antagonist SB-612111

[(-)-cis-1-Methyl-7-[[4-2，6-dichlorophenyl)piperidin-1-yl]methyl]-6，7，8，9-tetrahydro-5H-benzocyclohepten-5-ol]：In Vitro Studies”JPET(2007)321：961 967.

contents of the experiment

Orphanin saline, morphine, saline, nociceptin receptor antagonist (SB-612111).

Test protocol/flow diagram

(A) 6 rats per test group were lightly anaesthetised with polyurethane (1.0g/kg, i.p., with minimal active titration in the absence of irritation) (Sherman and Loomis et al, 1996). The left and right cheeks and the right hind limb were blackened with ink to distribute the heat evenly.

(B)200-300G male SD rats were tested in varying degrees of thermal pain intensity A-delta vs. C-fiber on the left, right cheek and right hind limb (Yeomans et al, 1996). In the C-fiber experiment, the heat intensity was adjusted by varying the power supply voltage of the focused projector until the retraction latency was between 9 and 13 seconds. The intensity (volts) applied to each animal was recorded. To reduce potential tissue damage, a 20 second latency period served as a node to stop stimulation. Rats that did not respond to a supply voltage of 55 volts were not included in the study.

For the a-delta fiber experiment, the heat intensity was adjusted until the retraction occurred at a latency of 2 to 3 seconds. The intensity (volts) applied to each animal was recorded. To reduce potential tissue damage, the 6 second heat generation time served as a node to stop stimulation. Rats that did not respond to a supply voltage of 85 volts were not included in the study.

(C) Rats were treated as follows:

50 μ L of the analgesic antagonist injected subcutaneously ((SB-612111, 10.0mg/kg) on the left and right cheeks and right hind limb.

Intravenous 50 microliter of saline

The experimental animals were treated by eye-covering.

(D) After 45 minutes, 48. mu.L of the analgesic (50 mg/ml) was titrated into nasal rats (Frey Method; Thorneet al, 2004).

(E) The withdrawal latency for radiant heat stimulation of the left and right cheeks was measured after the withdrawal latency for each hind limb response to radiant heat stimulation was measured. Each animal was tested for its specific radiation amplitude (volts) established in the baseline experiment. Detection was performed immediately after dosing and at 15, 30, 45, 60, 90, 120, 150 and 180 minutes.

(F) Rats were euthanized by CO2 inhalation.

FIGS. 5 and 7 show the effect of the nociceptin receptor antagonist SB-612111 on the A-delta pain threshold test for reduction of thermal stimuli by nasal drops of nociceptin. Compared with pretreatment of rat subcutaneous injection of the nociceptin receptor antagonist and pretreatment of subcutaneous registration of physiological saline, the curative effect of rat nasal administration of the nociceptin is detected. In fig. 5, the Y-axis shows the withdrawal latency of the rat face (cheek) in response to thermal stimuli, the hindpaw selectively activating the system components of the a-delta fiber receptor at a rate that are subject to intense pain. The Y-axis in figure 7 shows the withdrawal latency of the rat hind limb in response to thermal stimuli, with the hind paw selectively activating the a-delta fiber receptor at a rate for the systemic components that are subject to intense pain. The X-axis indicates the time the analgesic effect is maintained after administration.

FIGS. 6 and 8 show the effect of the nociceptin receptor antagonist SB-612111 on the C-fiber pain threshold test for reduction of thermal stimuli induced by intranasal administration of nociceptin. Compared with pretreatment of rat subcutaneous injection of the nociceptin receptor antagonist and pretreatment of subcutaneous registration of physiological saline, the curative effect of rat nasal administration of the nociceptin is detected. In fig. 5, the Y-axis shows the cheek retraction latency in response to thermal stimulation, the rat cheek selectively activates the pain system components of the C-fiber receptor to causalgia at one rate. The Y-axis in fig. 7 shows paw withdrawal latency in response to thermal stimuli, with the rat hind limb selectively activating the system components of the C-fiber nociceptor experiencing burning pain at one rate. The X-axis indicates the time the analgesic effect is maintained after administration. The apparent suppression of the analgesic activity of the analgesic antagonist indicates that the analgesic effect of the analgesic is due to binding to NOP receptors.

The data from these experiments can be analyzed as follows: withdrawal latency is the average result of cheek and hindlimb reactions between test animals. Analysis of variance from replicate experiments was used to compare the latency response of the cheeks and hind limbs to A-delta or C fiber in the saline and antagonist groups.

Example 4:analgesic Activity of nasal drip N/OFQ on model of persistent facial pain in rodents

The paw formalin model is widely used to evaluate the effect of treatment on persistent pain in accepted and known analgesic treatments. We applied this model to the cheek to evaluate and compare the effects of nasal drop N/OFQ and intranasal oxytocin and 2 benchmarks: sumatriptan (i.p.) and morphine (i.n.). The results of this study showed that 0.1mg/kg N/OFQ caused significant inhibition of rubbing behavior (47.2%) compared to intranasal administration of either 0.1mg/kg oxytocin (47.8%) or 42mg/kg sumatriptan (43.8%). At a higher dose, 3mg/kg morphine produced a greater maximum inhibition (80.2%) (fig. 9). Therefore, nasal drops of N/OFQ produced significant analgesic effects on the persistent facial pain in rodent models. Importantly, no anesthesia or other significant side effects were observed in N/OFQ dosed rats.

Example 5:efficacy of N/OFQ in incision pain model

To demonstrate the utility of N/OFQ in post-operative pain, we evaluated the behavior of anti-allodynia in paw-incision model rats following intranasal administration of N/OFQ (Brennan, T.J., Vandermeulen, E.P. & Gebhart, G.F.Pan 64, 493-501 (1996)). N/OFQ (5.0mg/kg) was intranasally administered 24 hours after a small incision was made on the plantar surface of the hind limb of the rat. The effect of intranasal N/OFQ was compared to intravenous N/OFQ.

The plantar surface of the right hind limb was surgically prepared with provodine-iodine and the rats were anesthetized with 2.5% isoflurane. A 5 mm incision was made through the skin and under the bandage on the plantar surface, near the interface of the hairy/hairless skin. Thereafter, the wound was closed with 5-0 silk using 2 simple interrupted stitches. The wound area is then followed by the test animal being returned to the mouse cage (fed alone) after treatment with three times the antibiotic ointment. Retraction thresholds to mechanical stimulation were determined using standard von Frey catheters (touch test; Stoelting Co., Wood Dale, IL, USA) methods from before right hind limb surgery to 24 hours post surgery.

For experimental purposes, rats were separated by a grid (10 × 10 cm) and allowed to habituate for 15 minutes. The test was carried out using the Dixon top-down method (Dixon, W.J.Staircase bioassay: the up-and-down method.Neurosci Biobehav Rev15, 47-50(1991)) to determine the 50% withdrawal threshold (Chaplan, S.R., Bach, F.W., Pogrel, J.W., Chung, J.M. & Yaksh, T.L.quantitative assessment of tactilendolynia in the rat.J.Neurosci Methods 53, 55-63 (1994)).

At the start of the test, all rats had 4.31 filaments (2.0 grams). The plantar surface incision of the right hind limb was stimulated with von Frey fibrils, and the presence of withdrawal (withdrawal) was observed, with the lowest (withdrawal) or highest (no withdrawal) being detected. Testing in this manner was continued until 4 more reactions (regardless of type) were initiated after the first pullback. The 50% threshold was calculated based on the type of reaction and the final test filament. on Freymon surfaces (3.61 to 5.18), covering the range of bending forces (0.4-15 g) and diameters (178-483 microns) can give forces in the range of 3.92-147 mN.

The analgesic effect can also be expressed as the maximum possible efficacy (% MPE) calculated from the 50% threshold (response to treatment) as follows:

% MPE [ (therapeutic response-post-cut baseline value) ]

(upper limit reaction-standard value after incision) ] × 100%;

wherein the upper limit reaction is 15.0. The dose response table is shown in figure 10.

Similar to the results of the thermal stimulation experiments, nasal administration of 5.0mg/kg N/OFQ produced a stronger anti-allodynic effect (FIG. 11), with a slight delay in onset (maximum efficacy was reached within 60 minutes). The same dose of iv was observed that N/OFQ (fig. 12) did not cause reversal of postoperative allodynia. These results provide strong evidence that nasal administration, rather than systemic administration of N/OFQ, is effective for post-operative pain. This dichotomy again demonstrates the efficacy of nasal administration of N/OFQ mediated by direct absorption through the central nervous system.

Example 6:N/OFQ efficacy in cheek incision pain model

FIG. 13 shows the effect of N/OFQ (5mg/kg, i.n) on postoperative static punctate allodynia caused by cheek incisions. A5 mm incision (including underlying muscle) was made percutaneously over the shaved cheek and 5.0mg/kg N/OFQ was nasally administered 24 hours later. Surgery was performed under 2.5% isoflurane anesthesia and the wound was closed with 1 simple interrupted suture 5-0 wire. The 50% withdrawal threshold was measured after every 30 minutes post-dose using von Freyflameraments (touch test; Stoelting Co., Wood Dale, IL, USA) using Dixon top-down technique. The model was modified from the hindlimb plantar incision model (Brennan, T.J., Vandermeulen, E.P. & Gebhart, G.F. (1996) Pain 64; 493-. The Von Frey monofilaments used in cheek testing cover the range of flexural force (0.0081.4 g) and diameter (64254 um) that can be given a force in the range of 0.07813.72 mN. The analgesic effect can also be expressed as the maximum possible efficacy (% MPE) calculated from the 50% threshold (response to treatment) as follows:

% MPE [ (therapeutic response-post-cut baseline value) ]

(upper limit reaction-standard value after incision) ] × 100%;

wherein the upper limit reaction is 1.5. The dose response table is shown in figure 14.

Example 7:retained nerve injury model

The analgesic activity of N/OFQ was assessed using a standard rat nerve injury model, the reserved nerve injury (SNI) model, (Decosterd, I. & Woolf, C.J. pain 87: 149-58 (2000)). Two of the three branches of the sciatic nerve of one leg were surgically allergic to the plantar surfaces of the ipsilateral hind limbs. This model produces many neuropathic pain features such as marked hyperalgesia, but associated spinal nerve damage, no signal stiffness, and sustained pain.

The method comprises the following steps:

surgery was performed under isoflurane (3.5%) anesthesia. Once deep anesthesia was achieved, the outer surface skin of the thigh was shaved and treated with providine-iodine solution. The skin was incised and an incision was made to expose the sciatic nerve and three terminal branches: fibula, common fibula and tibial ganglia. The tibial ganglia and common fibula were cut and ligated with 5.0 silk to leave the calf nerve intact, but the 2-4 mm distal nerve end was removed. The nursing is well done, any contact is avoided, or the calf nerve is stretched. The muscle and skin are wrapped and sealed by double layers, and the wound is treated by antibiotic ointment. After surgery, 0.05mg/kg buprenorphine IM was used as postoperative analgesia. To assess the effect of nerve injury on hyperalgesia, mechanical pain (touch test; Stoelting co., Wood Dale, IL, USA) was measured using von Frey filaments, applied to the plantar ipsilateral hind limbs and the effect before and after injury, before and after nasal drip N/OFQ was assessed. And detecting a 50% withdrawal threshold value by using a Dixon top-to-bottom technology. The model was modified from the hindlimb plantar incision model (Brennan, T.J., Vandermeulen, E.P. & Gebhart, G.F. (1996) Pain 64; 493-. The Von Frey monofilaments (monofil filaments) used in the test cover the range of bending forces (0.0081.4 g) and diameters (64254 um) that can be given a force in the range of 0.07813.72 mN.

As a result:

nerve injury to the ipsilateral paw injured within 6 days of surgery produced significant allodynia. N/OFQ (5.0mg/kg) was administered nasally 21 days after nerve injury, and the ipsilateral paw nerves produced a strong analgesic effect throughout the 3 hour test (FIG. 13). These effects are no longer evident 24 hours after administration, i.e. on the following day. Despite the small samples (Vehicle: N4 & N/OFQ: N6), there is still a clear trend towards efficacy. These results provide strong evidence that nasal drops of N/OFQ are effective in treating neuropathic pain, except at the original pain site.

Example 8:therapeutic effect of N/OPQ on intestinal tract

One of the major complications associated with pain management is constipation, with administration of opioid analgesics, such as morphine. Constipation increased over the time of intestinal transit in rats. To determine whether nasal administration of N/OPQ, which has a similar strong analgesic effect as opioids, would also cause constipation, the effect of nasal administration of N/OFQ on intestinal transit time was evaluated. Rats were tested for Intestinal Transit Time (ITT) after N/OFQ (5.0mg/kg) pretreatment and given charcoal powder (12.5% activated carbon activated with a 12.5% gum arabic suspension) (Tavani, A., Petrillo, P., La Regina, A. & Sbacchi, M.role of peripheral mu, delta and kappa opioid receptors in-induced inhibition of synergistic therapeutic transit in rates.JPharmacol Exp Ther 254, 91-97 (1990)).

The effects of intraperitoneal (3.0mg/kg) or nasal (4.0mg/kg) morphine were compared. Briefly, rats were given ad libitum feed and water overnight, respectively, and the test compound was administered the next morning. After 20 to 30 minutes, the rats were gavaged with charcoal suspension (2 ml) and after 30 minutes, euthanized by inhalation of carbon dioxide. The stomach and small intestine were immediately excised, and the total length of the small intestine and the distance traveled by charcoal were recorded. Calculated by (transit)% multiplied by 100% of the total length of small intestine passed divided by the distance traveled by charcoal. For cross-over study comparisons, the effect of the complexes can also be expressed as% transit of vehicle treated group (100% - ((mean vehicle% traversed distance-drug% traversed distance)/mean vehicle% traversed distance))%. Although morphine produced some slowing of transit (drop), no effect was detected in N/OFQ-pretreated rats regardless of the route of administration (figure 14). These data provide strong evidence that nasal drops of N/OFQ do not produce clinically significant constipation despite producing a robust analgesic effect similar to morphine.

Example 9:preliminary evaluation of behavior after nasal drip N/OFQ

It is well known that there are sometimes profound behavioral toxicological effects (i.e., sedation) as clinically useful potent analgesics, and it is important to determine whether N/OFQ is equally burdened. Therefore, preliminary behavioral toxicology screening of nasal drops N/OFQ was examined. Briefly, 4 rats were nasally dosed with 25mg/kgN/OFQ and returned to their cages (housed individually) and allowed to recover from anesthesia. Every 20 minutes, after one hour, every 1 hour, and any abnormalities noted.

24 hours after N/OFQ administration, mice were again observed for any delayed toxic response. Generally, the determination of whether a rat behaves abnormally is subjectively determined, and there is one of the following behaviors: tonic/clonic movements, ataxia, stereotypy, vocalization, lacrimation, salivation, hair uprighting, ptosis, and abnormal breathing. Furthermore, treatment of the animal or proximity or handling of the animal if any abnormal response is elicited is considered a stimulated response. No abnormal response was observed when the acute toxic dose was increased 5-fold over the thermally stimulated and incisional model (5mg/kg) effective dose.

While the foregoing invention has been described in some detail by way of illustration and example for purposes of clarity of understanding, it will be readily apparent to those of ordinary skill in the art that certain changes and modifications may be made without departing from the invention as defined by the appended claims. Therefore, the description and examples should not be construed as limiting the scope of this patent.

It should be emphasized that the singular forms "a," "an," and "the" include plural referents unless otherwise specified. Furthermore, as used herein, the terms "includes," "including," and its derivatives are intended to be inclusive, i.e., equivalent to the term "comprising" and its corresponding homologous meaning.

All cited patents, patent applications, documents, articles are incorporated by reference in their entirety.

Claims (36)

1. A method of treating pain comprising administering to an individual in need thereof an effective dose of an NOP agonist via craniofacial mucosal administration.

2. The method of claim 1, wherein the treatment comprises treatment of trigeminal neuralgia, somatic pain, neuropathic pain, or visceral pain.

3. The method of claim 1, wherein the treatment comprises treatment of acute or chronic pain.

4. The method of any one of claims 1-3, wherein the pain is craniofacial pain or headache pain.

5. The method of claim 4, wherein the craniofacial pain or headache is pain resulting from a temporomandibular joint disorder (TMJ), migraine or trigeminal neuralgia.

6. The method of any of claims 1-5, wherein craniofacial mucosal administration comprises nasal administration, buccal administration, sublingual administration, or conjunctival administration.

7. The method of claim 6, wherein mucosal administration is nasal administration.

8. The method of any one of claims 1-7, wherein the NOP agonist is a NOP agonist peptide.

9. The method of any one of claims 1-8, wherein the NOP agonist is N/OFQ.

10. The method of any one of claims 1-9, further comprising administering to the subject in need thereof at least one or at least two additional active agents, wherein the additional active agents are administered prior to, after, or simultaneously with the administration of the NOP agonist.

11. The method of any one of claims 10, wherein the additional active agent is selected from the group consisting of: non-opioid analgesic peptides, opioid or opioid-like peptides or analogs thereof, NMDA receptor antagonists, sodium channel blockers, calcium channel blockers, adrenergic receptor antagonists, GABAergic agonists, glycine receptor agonists, cholinergic agonists, adrenergic receptor agonists, e.g., epinephrine, anticonvulsants, Rho kinase inhibitors, PKC inhibitors, P38-MAP kinase inhibitors, ATP receptor blockers, endothelin receptor blockers, pro-inflammatory cytokines, chemokines, interleukins, and tumor necrosis factor receptor blockers, pro-inflammatory cytokines, tricyclic antidepressants, serotonin antagonists, serotonin receptor agonists non-steroidal anti-inflammatory drugs, NSAIDs and COXIBs, acetaminophen, analgesic peptides, toxins, TRP channel agonists and antagonists, canabanoids antagonists, analgesic neurotransmitter receptors CGRP 1and CGRP2, nociceptin neurotransmitter receptor NK1 receptor antagonists, antagonists algesin NK2 neurotransmitter receptors, nociceptive peptide neurotransmitter receptor Y1-5, nociceptive peptide neurotransmitter receptors VPAC2, VPAC1and PAC1, algesin neurotransmitter receptor GAL1-3 or antagonist GalR1, 3, agonists or vasopressin receptor antagonists, corticoid releasing factor (CRH), growth hormone releasing factor (GHRH), luteinizing hormone releasing factor (LHRH), somatostatin growth hormone release inhibitor, thyroid hormone releasing factor (TRH), brain-derived neurotrophic factor (BDNF) of glial cell-derived neurotrophic factor (GDNF), Nerve Growth Factor (NGF), neurotrophic factor-3 (NT-3), pancreatic polypeptide, polytyrosine, glycomacropeptide 1(GLP-1), histidine isoleucine peptide (phenanthrene), Pituitary Adenylate Cyclase Activating Peptide (PACAP), brain natriuretic peptide (Naja), CCK, islet amyloid polypeptide (IAPP) or amylin, Melanin Concentrating Hormone (MCH), melanocortin (ACTH, alpha-MSH and others), neuropeptide FF (F8Fa), neurotensin, parathyroid hormone-related protein, calcitonin, agouti gene-related protein (AGRP), Cocaine and Amphetamine Regulated Transcript (CART)/peptide, 5-HT-moduline, hypothalamic hormone/orexin, oxytocin, ocistatin, urocortin, secretory neuron, prolactin release peptide and derivatives and analogs thereof.

12. The method of any one of claims 1-9, wherein the NOP agonist is administered in a pharmaceutical composition.

13. The method of claim 12, wherein the pharmaceutical composition further comprises at least one or at least two additional active agents.

14. The method of claim 12, wherein the pharmaceutical composition further comprises one or more additional active agents selected from the group consisting of: non-opioid analgesic peptides, opioid or opioid-like peptides or analogs thereof, NMDA receptor antagonists, sodium channel blockers, calcium channel blockers, adrenergic receptor antagonists, GABAergic agonists, glycine receptor agonists, cholinergic agonists, adrenergic receptor agonists, such as epinephrine, anticonvulsants, Rho kinase inhibitors, PKC inhibitors, P38-MAP kinase inhibitors, ATP receptor blockers, endothelin receptor blockers, pro-inflammatory cytokines, chemokines, interleukins and tumor necrosis factor receptor blockers, pro-inflammatory cytokines, tricyclic antidepressants, serotonin antagonists, serotonin receptor agonists non-steroidal anti-inflammatory drugs, NSAIDs and COXIBs, acetaminophen, analgesic peptides, toxins, TRP channel agonists and antagonists, canabanoids antagonists, analgesic neurotransmitter receptors CGRP 1and CGRP2, nociceptin neurotransmitter receptors NK1 receptor antagonists, antagonists algesin NK2 neurotransmitter receptors, nociceptive peptide neurotransmitter receptor Y1-5, nociceptive peptide neurotransmitter receptors VPAC2, VPAC1and PAC1, algesin neurotransmitter receptor GAL1-3 or antagonist GalR1, 3, agonists or vasopressin receptor antagonists, corticoid releasing factor (CRH), growth hormone releasing factor (GHRH), luteinizing hormone releasing factor (LHRH), somatostatin growth hormone release inhibitor, thyroid hormone releasing factor (TRH), brain-derived neurotrophic factor (BDNF) of glial cell-derived neurotrophic factor (GDNF), Nerve Growth Factor (NGF), neurotrophic factor-3 (NT-3), pancreatic polypeptide, polytyrosine, glycomacropeptide 1(GLP-1), histidine isoleucine peptide (phenanthrene), Pituitary Adenylate Cyclase Activating Peptide (PACAP), brain natriuretic peptide (Nalcin), CCK, islet amyloid polypeptide (IAPP) or amylin, Melanin Concentrating Hormone (MCH), melanocortins (ACTH, alpha-MSH and others), neuropeptide FF (F8Fa), neurotensin, parathyroid hormone-related protein, calcitonin, agouti gene-related protein (AGRP), Cocaine and Amphetamine Regulated Transcript (CART)/peptide, 5-HT-moduline, hypothalamic hormone/orexin, oxytocin, ocistatin, urocortin, secretin, prolactin release peptide and derivatives and analogs thereof.

15. The method according to any one of claims 12-14, wherein the pharmaceutical composition further comprises one or more pharmaceutically acceptable excipients, adjuvants, diluents or stabilizers.

16. The method according to any one of claims 12-15, wherein the pharmaceutical composition further comprises at least one protease inhibitor and/or at least one absorption enhancer.

17. The method of any one of claims 12-16, wherein the NOP agonist is a NOP agonist peptide.

18. The method of any one of claims 12-17 wherein the NOP agonist is N/OFQ.

19. A method of treating or preventing migraine pain comprising administering to an individual in need thereof an effective dose of an NOP agonist, wherein the NOP agonist is administered nasally.

20. The method of claim 19 wherein the NOP agonist is N/OFQ.

21. The method of claim 9, wherein the unit dose of N/OFQ administered is from about 0.2mg to about 5000 mg.

22. A method for treating migraine comprising administering to an individual in need thereof an effective amount of N/OFQ by nasal administration.

23. The method of claim 22, wherein treating comprises treating one or more symptoms associated with migraine selected from the group consisting of nausea, photophobia and phonophobia.

24. The method of claim 22 or 23, wherein the migraine is migraine without aura, migraine with aura, or migraine with aura but without headache.

25. The method of any one of claims 1-21, wherein administration results in a 30% or greater reduction in VAS pain score.

26. A kit comprising an NOP agonist and instructions for treatment of pain, wherein the instructions comprise instructions for administration of the NOP agonist via the craniofacial mucosal route.

27. The kit of claim 26, further comprising a drug delivery device.

28. The kit of any one of claims 26 or 27 wherein the NOP agonist is N/OFQ.

Use of a NOP agonist in the manufacture of a medicament for administration via craniofacial mucosa for the treatment of pain.

30. The use according to claim 29, wherein craniofacial mucosal administration is nasal administration.

31. Use according to claim 29 or 30 wherein the NOP agonist is a NOP agonist peptide.

32. The use of any one of claims 29-31 wherein the NOP agonist is N/OFQ.

33. An NOP agonist for use in the treatment of pain by craniofacial mucosal administration.

34. An NOP agonist for use in the treatment of pain by nasal administration.

35. The NOP agonist of claim 33 or 34 wherein the NOP agonist is a NOP agonist peptide.

36. The NOP agonist of any one of claims 33 to 35 wherein the NOP agonist is N/OFQ.