[go: up one dir, main page]

WO1990013293A1 - Compositions de cytochalasine et procedes therapeutiques - Google Patents

Compositions de cytochalasine et procedes therapeutiques Download PDF

Info

Publication number
WO1990013293A1
WO1990013293A1 PCT/US1990/002342 US9002342W WO9013293A1 WO 1990013293 A1 WO1990013293 A1 WO 1990013293A1 US 9002342 W US9002342 W US 9002342W WO 9013293 A1 WO9013293 A1 WO 9013293A1
Authority
WO
WIPO (PCT)
Prior art keywords
cytochalasin
cytochalasins
administration
group
immunosuppression
Prior art date
Application number
PCT/US1990/002342
Other languages
English (en)
Inventor
Thomas P. Fondy
Dennis Bogyo
Original Assignee
Syracuse University
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Syracuse University filed Critical Syracuse University
Publication of WO1990013293A1 publication Critical patent/WO1990013293A1/fr

Links

Classifications

    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D491/00—Heterocyclic compounds containing in the condensed ring system both one or more rings having oxygen atoms as the only ring hetero atoms and one or more rings having nitrogen atoms as the only ring hetero atoms, not provided for by groups C07D451/00 - C07D459/00, C07D463/00, C07D477/00 or C07D489/00
    • C07D491/02—Heterocyclic compounds containing in the condensed ring system both one or more rings having oxygen atoms as the only ring hetero atoms and one or more rings having nitrogen atoms as the only ring hetero atoms, not provided for by groups C07D451/00 - C07D459/00, C07D463/00, C07D477/00 or C07D489/00 in which the condensed system contains two hetero rings
    • C07D491/10—Spiro-condensed systems
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/40—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having five-membered rings with one nitrogen as the only ring hetero atom, e.g. sulpiride, succinimide, tolmetin, buflomedil
    • A61K31/403—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having five-membered rings with one nitrogen as the only ring hetero atom, e.g. sulpiride, succinimide, tolmetin, buflomedil condensed with carbocyclic rings, e.g. carbazole
    • A61K31/4035—Isoindoles, e.g. phthalimide
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents

Definitions

  • the present invention is directed to naturally occurring and synthetic cytochalasin compositions and therapeutic treatments utilizing these compositions. More specifically, the present invention relates to certain synthetic analogues of Cytochalasin B (CB) and sustained release formulations containing a cyctochalasin, for example, cytochalasin B and other natural cytochalasins, for example, Cytochalasin D, E, H or J, among others, and one or more of its synthetic analogues.
  • CB Cytochalasin B
  • This invention also relates to the surprising discovery that the administration of cytochalasins including CB produces transient immunosuppression which is controllable by dose or route of administration and is reversible spontaneously or with the use of IL-2.
  • a therapeutic regimen of cytochalasins may be used to treat the undesirable hyperimmunity of transplant patients and patients with autoimmune disease.
  • anti-tumor therapy utilizing CB and other cytochalasins, and optionally antineoplastic agents other than cytochalasins may be significantly enhanced by combining the administration of these agents with effective amounts of IL-2 or other lymphokines for reversing the immunosuppression produced during administration of cytochalasins or cytochalasins with other antineoplastic agents.
  • This invention also relates to sustained release formulations utilizing liposomes or microcapsules which are effective for delivering high concentrations of cytochalasins and optionally, additional antineoplastic agents to the active site of the tumor without producing undesirable immunosuppression.
  • the cytochalasins, membrane- and transport-acting compounds include the congeneric cytochalasins A-M as well as the semi-synthetic derivatives 7,20- di-O-acetyl-cytochalasin B, 7-mono-0-acetyl-cytochalasin B, 21,22-dihydro-cytochalasin B and 21,22-dihydro-cytochalasin A, together with the related chaetoglobosins, constitute a class of more than 24 structurally and functionally related alkaloid metabolites produced by molds (Yahara et al., 1982, J.
  • cytochalasin B CB
  • CB cytochalasin B
  • the cytochalasins are also known to alter microfilament morphology (Schliwa, 1982, J. Cell Biol., 92:79-91), thereby affecting the cellular functions that depend upon microfilament biochemistry.
  • Some of the cellular processes sensitive to the cytochalasins are phagocytosis, pinocytosis, cytokinesis, secretion, and exocytosis, as well as functions requiring movement and/or intercellular adherence including intracellular organelle movement and intercellular transport, cell motility, transport across tissue barriers, and a variety of immunological responses.
  • the cytochalasins have only recently been shown to possess substantial activity against tumor growth and metastasis.
  • cytochalasins and especially, CB inhibit the growth and metastases of tumors, extend the tumor latency period, and extend host survival in a murine carcinoma and murine melanoma model system (Bousquet, et al., submitted for publication).
  • Q Cytochalasins may function as chemotherapeutic amplifiers of the activity of known anti-tumor agents by virtue of the effects on cytoskeletal and plasma membrane functions.
  • cytochalasins have been shown to affect leukocyte-mediated functions in vitro, the differentiation of cytolytic T lymphocytes and also to affect cell motility, cell adherence, phagocytosis and secretion
  • Certain of these currently used agents also find limited use in treating autoimmune diseases.
  • the therapy suffers from the same 5 limitations which occur when immunosuppressive agents are used to treat transplant or tissue graft rejection.
  • the search therefore, continues for more effective immunosuppressive agents that are effective at controlling a heightened immune response (hyperimmunity) in patients with transplanted organs, grafted tissue Q or an autoimmune disease without affecting the ability of the patient's system to fight off infection.
  • the present invention also relates to the surprising discovery that the cytochalasins, especially CB, exhibit significant transient, readily reversible immunosuppression in vivo.
  • Cytochalalasins may become drugs of choice, either alone or in combination with known antineoplastic agents for the treatment of tumors. CB, however, after administration, metabolizes quickly to the more highly toxic cytochalasin A (CA), as well as other metabolites.
  • CA cytochalasin A
  • Past studies have suggested that CA is highly reactive with thiol groups, a fact which might explain the high toxicity associated with its use. It is believed that the metabolic oxidation of the 20-hydroxyl group of CB to the 20 keto group in CA to form the highly reactive enedione system, a chemical system which is thought to be highly reactive with thiols in enzymes and other proteins, may be primarily responsible for the high toxicity associated with the administration of CA.
  • the cytochalasins exhibit a number of pharmacological effects, many of which may be controlled by the amount and route of administration. These pharmacological effects may also be 0 controlled by formulations utilizing liposome and microencapsulation techniques, or alternatively, by the administration of cytochalasins with, for example, lymphokines.
  • Liposomes are completely closed lipid bilayer membranes which g contain entrapped aqueous volume. Liposomes are vesicles which may be unilamellar (single membrane) or multilammelar (onion-like structures characterized by multiple membrane bilayers, each separated from the next by an aqueous layer).
  • the bilayer is composed of two lipid monolayers having a hydrophobic "tail" region Q and a hydrophilic "head” region. In the membrane bilayer, the hydrophobic (nonpolar) "tails" of the lipid monolayers orient toward the center of the bilayer, whereas the hydrophilic (polar) "heads” orient toward the aqueous phase.
  • the basic structure of liposomes may be made by a variety of techniques known in the art.
  • the invention of the present application also relates to the discovery that the use of interleukin-2 (IL-2) in combination with the cytochalasins and optionally, other antineoplastic agents, eliminates the immunosuppressive effects of the cytochalasins.
  • Interleukin-2 a lymphokine which is produced by normal peripheral blood lymphocytes and induces proliferation of antigen or mitogen stimulated T cells after exposure to plant lectins, antigens or other stimuli, was first described by Morgan, D. A., et al.., Science, 193, 1007 (1976). 11-2, in addition to its ability to induce proliferation of stimulated T lymphocytes, also modulates a number of functions of immunocytes in vivo.
  • IL-2 is one of several lymphocyte-produced messenger regulatory molecules which mediate immunocyte interactions and functions.
  • Figure 1 shows the maximum tolerated dosage (MTD) for CB administered to B6D2F1, CD2F1 or C57B1/6 via intraperitoneal (IP), sub-cutaneous (SC) and intravenous (IV) routes of administration using different vehicles.
  • IP intraperitoneal
  • SC sub-cutaneous
  • IV intravenous
  • Figure 2 shows the effect of varying doses of CB administered to mice 19 hours before splenectomy on immunosuppression.
  • Figure 3 shows the effect of varying doses of CB administered to mice 3 hours before splenectomy on immunosuppression.
  • Figure 4 shows the effect of the IP administration of 50 mg/kg on the allogeneic rejection response in vivo.
  • Figure 5 shows an absence of suppression of specific cytotoxicity nine days after groups of mice were treated in vivo with 50 mg/kg CB at any time before or after tumor challenge when compared to vehicle-treated controls.
  • Figures 6, 7 and 8 shows the reversal of immunosuppression produced by adding IL-2 after mice were treated with 50 mg/kg of CB at 19 hours, 3 hours and 72 hours before splenectomy.
  • Figure 9 compares the cytotoxicity exhibited by spleen cells which are washed prior to treatment with rIl-2 with the cytotoxicity exhibited by unwashed spleen cells treated with rIl-2.
  • Figure 10 compares the immunosuppression produced by high doses of CB administered in microcapsules with the immunosuppression produced by CB at doses varying between 2 mg/kg to 50 mg/kg administered IP.
  • Figure 11 shows the effect of treatment with CB at various times before and after tumor challenge on tumor-induced splenic enlargement.
  • Figure 12 shows the effect of treatment with CB at various times before and after tumor challenge on tumor-induced splenic cellularity.
  • the present invention relates to pharmaceutical compositions and therapeutic methods for treating mammals, especially humans, with cytochalasins to reversibly suppress the heightened immune response (hyperimmunity) in autoimmune disease states or following organ transplants or tissue grafts.
  • an immunosuppressive dose comprising no less than about one fifth the maximum tolerated dosage of a cytochalasin is administered intravenously, intramuscularly, subcutaneously or orally to a patient suffering from hyperimmunity.
  • the present invention also relates to intramuscular, intravenous, subcutaneous and oral dosages of cytochalasins which may be useful for inhibiting the growth and metastasis of tumors without producing concomitant immunosuppressive effects.
  • cytochalasins comprising no greater than about one fifth the maximum tolerated dosage for a particular route of administration are formulated and administered alone or in combination with other antineoplastic agents for the inhibition of the growth of tumors.
  • the present invention also relates to the discovery that IL-2 as well as other lymphokines may be used to eliminate the immunosuppressive effects produced by the cytochalasins.
  • pharmaceutical compositions and therapeutic methods employing cytochalasins and IL-2 in combination are presented.
  • an additional antineoplastic agent other than a cytochalasin may be included for administration with the cytochalasin and IL-2.
  • the present invention also relates to pharmaceutical compositions and therapeutic methods utilizing cytochalasins in liposomes or microcapsules to provide sustained release formulations of cytochalasin which deliver large antineoplastic doses of cytochalasin to the site of a tumor without producing concomitant immunosuppressive effects.
  • a surprising discovery of the present invention is that concentrations of cytochalasin up to about three times the maximum therapeutic dosage may be administered in vivo in liposomes or microcapsules without producing the immunosuppressive effects which exist when cytochalasins are not administered in liposomes or microcapsules.
  • the present invention further relates to semi-synthetic cytochalasin analogues, pharmaceutical compositions containing these analogues and therapeutic methods utilizing these analogues which do not readily metabolize to the enedione system of cytachalasin A (CA).
  • CA cytachalasin A
  • Cytochalasins synthetic or naturally occurring mold-derived microfilament and membrane-acting compounds.
  • DMSO dimethyl sulfoxide
  • Tw - Tween polyoxyethylene sorbitan monoalkyl ethers
  • surface-acting agents VET - vesicles made by an extrusion technique
  • the present invention relates to a method for producing transient immunosuppression in mammals using cytochalasins. More specifically, it has been discovered that the cytochalasins may be used as transient immunosuppressive agents for treating hyperimmunity associated with organ transplants, tissue grafts and autoimmune disease, for example rheumatoid arthritis, lupus, autoimmune diabetes, autoimmune thyroiditis, autoimmune hepatitis.
  • a cytochalasin is administered to a mammal in an amount within the range of about one fifth the maximum tolerated dose (MTD) to about the MTD for a particular route of administration, preferably about one half to about the MTD of cytochalasin, and most preferably about the MTD.
  • MTD maximum tolerated dose
  • cytochalasins including cytochalasins A-M as well as the semi-synthetic derivatives of cytochalasin, including 21,22-dihydro-cytochalasin B, 20- deoxy-cytochalasin B, 20-deoxy-21,22-dihydrocytochalasin B, 20- deoxy-20-fluoro-cytochalasin B, 20-deoxy-20-fluoro-21, 22- dihydrocytochalasin B and- 21,22-dihydro-cytochalasin A, among , others, may be used in the immunosuppressive method of the present invention.
  • cytochalasins differ in their ability to inhibit microfilament formation, phagocytosis, cytokinesis, secretion and exocytosis, and consequently, in their ability to produce immunosuppression.
  • route of administration and the pharmacokinetics of the cytochalasin derivative also play an important role in determining the extent of immunosuppression.
  • immunosuppressive dosages of natural and semi-synthetic cytochalasins fall within the range of about one fifth to about the MTD for a particular route of administration.
  • the daily dosage of cytochalasin effective for producing transient immunosuppression will generally range from about 0.04 mg/kg to about 150 mg/kg and preferably from about 1.0 mg/kg to about 150 mg/kg.
  • concentration of cytochalasin used will be varied according to the route of administration and activity of the cytochalasin.
  • the amount of cytochalasin used generally ranges from about 2 mg/kg to o about 150mg/kg and preferably 5 mg/kg to about 150 mg/kg.
  • the amount of cytochalasin-used will generally range from about 0.1 mg/kg to about 20 mg/kg and preferably 0.25 mg/kg to about 20 mg/kg.
  • the amount of cytochalasin used will generally range from about 1 mg/kg to about 50 mg/kg and preferably about 2.5 mg/kg to about 50 mg/kg.
  • effective concentrations of CE and CD useful in the present invention will generally be much smaller (about one tenth) than the concentration of CB used because of the difference in potency.
  • the cytochalasins are administered in an amount equal to about the MTD for a particular route of administration.
  • the table presented in figure 1 presents representative MTD for a number of different CB containing vehicles administered to mice IP, SC or IV.
  • typical pharmaceutical formulation solvents and delivery suspensions may be utilized.
  • the formulations are generally administered in admixture with a pharmaceutical carrier selected with regard to the intended route of administration and standard ' pharmaceutical practice.
  • the cytochalasins may be used in the form of a sterile, pyrogen-free aqueous solution which may contain other solutes, for example, salts or sugars such as glucose to make the solution isotonic. Any suspension or solvent which is pharmaceutically compatible may be used in the present invention.
  • the type of suspension or solvent used in the parenteral formulations of the present invention may affect the absorptivity of the cytochalasin. Modifying the formulations to maximize the immunosuppressive effect of the cytochalasins is well within the skill of one of ordinary skill in the formulation arts.
  • Preferred suspensions for parenteral administration include cytochalasins in CMC 2%/Tween-20 1%.
  • Preferred solutions include cytochalasins in ethanol/saline (1:2) and DMSO.
  • the immunosuppressive effect of the cytochalasin used may be affected by the site as well as the route of administration.
  • the transient, reversible immunosuppressive effect of the cytochalasins is dependent on timing and the amount of cytochalasin to reach the site of activity.
  • Sites of activity for cytochalasin immunosuppression may include the spleen, bone marrow, lymph nodes, thymus or a transplanted organ or graft site.
  • One of ordinary skill in the art will know to vary the amount, route and site of administration to maximize the concentration of cytochalasin at the site of immunosuppressive activity.
  • cytochalasins When intravenous administration of cytochalasins is contemplated, care must be taken to choose the site of administration to maximize localization of cytochalasin at the site of immunosuppression within a short time frame, i.e., about one to twelve hours after- administration, and preferably, one tq about three hours after administration.
  • Immunosuppressive amounts of cytochalasin may also be administered orally in gelatin capsules, powders, syrups, elixirs, aqueous solutions, suspensions and the like.
  • Oral dosage forms are preferably administered as immediate release oral products.
  • An immediate release oral product is one that releases the active agent immediately after the product reaches the gut.
  • cytochalasins which are formulated as immediate release products may reach certain sites of immunosuppressive activity, for example, the spleen, as readily as the parenterally administered cytochalasins, thus promoting the maximum 0 immunosuppressive effect.
  • the orally administered products may be administered in combination with pharmaceutical carriers and diluents, for example, _ lactose, sodium citrate, salts of phosphoric acid, magnesium stearate, starch and talc.
  • a preferred route of oral administration is via immediate release soft gelatin capsules in which the cytochalasin is solubilized in a lower molecular weight polyethylene glycol, or another solvent, for example, DMSO, or mixtures thereof, o so as to maintain the cytochalasin in a soluble or near-soluble state in the capsule to maximize dissolution, absorptivity and blood concentration of the cytochalasin after the capsule dissolves in the GI tract.
  • the immunosuppression produced by cytochalasins is transient and readily reversible.
  • the immunosuppression produced by a bolus dose of cytochalasin generally increases or decreases as a function of the concentration of the cytochalasin at the site of immunosuppressive activity. Therefore, immunosuppression produced by cytochalasins may be carefully controlled with a therapeutic regimen designed to maximize the concentration of cytochalasin at the site of immunosuppression using cytochalasin pharmacokinetic data.
  • Treatment may vary such that, an immunosuppressive amount of a cytochalasin may be administered as a bolus dose once or twice daily via a parenteral route, or orally, once every six to twelve hours (qid or bid).
  • the cytochalasins may be administered at the onset of heightened immunity.
  • the therapeutic regimen chosen will vary as a function of the activity, route of administration and pharmacokinetics of the cytochalasin chosen.
  • the present invention also relates to antineoplastic dosage forms of cytochalasin and methods of treatment which inhibit the growth and spread of tumors without producing concomitant immunosuppressive effects.
  • a cytochalasin is administered to a patient in an amount equal to no greater than about one fifth of the MTD, and preferably no greater than about one tenth the MTD.
  • the type, route of administration and pharmacokinetics of the cytochalasin derivative used play an important role in determining the proper dosage.
  • cytochalasin dosages which inhibit the growth and metastasis of tumors but do not produce concomitant immunosuppression fall within the range of about 0.05 mg/kg to about 30 mg/kg.
  • the cytochalasins may be administered parentally or orally in the same manner and using the same solvents and other additives that are used in the immunosuppressive aspect of the present invention, except that lower dosages are to be used to avoid significant immunosuppression.
  • the cytochalasins in this aspect of the present invention may be formulated in combination with certain antineoplastic agents other than cytochalasins; however, it is preferred that these agents should not themselves produce significant immunosuppression unless additional therapy for eliminating immunosuppression is also used.
  • the present invention also relates to the surprising discovery that sustained release formulations comprising cytochalasins and liposomes and/or microcapsules in dosages significantly higher than those which produce immunosuppression may be administered to patients without causing appreciable immunosuppression.
  • Dosage forms of cytochalasin comprising at least about the maximum tolerated dosage, and preferably up to about three times the maximum tolerated dosage for a route of administration formulated in liposomes or microcapsules are useful for treating neoplasia without producing immunosuppression.
  • antineoplastic agents may be formulated along with the cytochalasin in the liposomes or microcapsules, it is preferred that only those neoplastic agents which exhibit an absence of immunosuppression should be formulated in combination with cytochalasin unless additional therapy for eliminating immunosuppression is also used.
  • Liposomes may be used as the sustained release delivery vehicle for the administration of the cytochalasins and optionally, an antineoplastic agent. Any of the techniques known in the art for making liposomes may be used in this invention. For example, the
  • Unilamellar vesicles may be produced using an extrusion apparatus by a method described in Cullis et al., PCT Publication No. WO 87/00238, published January 16, 1986, entitled "Extrusion Technique for Producing Unilamellar Vesicles” incorporated herein by reference. Vesicles made by this technique, called LUVETS, are 0 extruded under pressure through a membrane filter. Vesicles may also be made by an extrusion technique through a 200 nm filter; such vesicles are known as VET200s.
  • liposomes that may be used are those 5 characterized as having substantially equal lamellar solute distribution.
  • This class of liposomes is denominated as stable plurilamellar vesicles (SPLV) as defined in U.S. Patent No. 4,522,803 to Lenk, et al., monophasic vesicles as described in U.S.
  • SPLV plurilamellar vesicles
  • any of the methods for making liposomes can be used to 1Q encapsulate the semi-synthetic or naturally occurring cytochalasins.
  • Pharmaceutical compositions of these liposomal forms of the cytochalasins can be administered in vitro or in vivo as described hereinbelow.
  • the liposomes can be loaded with drug according to the procedures of Bally et al., PCT 0 Publication No. 86/01102, published February 27, 1986, and incorporated herein by reference.
  • This technique allows the loading of antineoplastic agents by creation of a transmembrane concentration gradient across the liposome membranes. This gradient is generated by a concentration gradient for one or more ionic species (e.g., Na+, C1-, -K+, Li+, or H+) across the liposome membranes.
  • these ionic gradients are pH (H+) gradients, which drive the uptake of the ionizable antineoplastic agent across the liposome membranes.
  • H+ pH
  • pharmaceutical formulations can be made which can be delivered in vitro or in vivo as described hereinbelow.
  • a bioactive agent such as a drug is entrapped in or associated with the liposome and then administered to the patient to be treated.
  • a bioactive agent such as a drug
  • U.S. Patent No. 3,993,754 Sears, U.S. Patent No. 4,145,410; Papahadjopoulos et al., U.S. Patent No. 4,235,871; Schneider, U.S. Patent No. 4,114,179; Lenk et al., U.S. Patent No. 4,522,803; and Fountain et al., U.S. Patent No. 4,588,578.
  • the semi- synthetic and naturally-occurring mold-derived cytochalasins may be entrapped in or associated with liposomes.
  • antineoplastic agents or agents which are administered to eliminate the immunosuppressive effects of the cytochalasins or the additional antineoplastic agents as described hereinabove can be co-entrapped in liposomes, or entrapped in liposomes co-administered with those liposomes containing the cytochalasins.
  • Such formulations of liposomes may be administered simultaneously or sequentially.
  • a preferred method of the present invention is to entrap up to three times the MTD of cytochalasin alone or in combination with an effective amount of an additional antineoplastic agent. This formulation is then administered to the patient as a sustained release form without producing substantial immunosuppression.
  • organic solvents may be used to suspend the lipids.
  • Suitable organic solvents for use in the present invention include those with a variety of polarities and dielectric properties, which solubilize the lipids, for example, chloroform, methanol, ethanol, dimethylsulfoxide (DMS0), methylene choloride, and solvent mixtures such as benzene:methanol (70:30), among others.
  • solvents are generally chosen on the basis of their biocompatability, low toxicity, and solubilization abilities.
  • Liposomes containing the pharmaceutical formulations including cytochalasins of the present invention may be used therapeutically in mammals, especially humans, in the treatment of neoplasms which require repeated administration.
  • the sustained release formulations utilize up to three times the MTD of cytochalasin. Such sustained release compositions are effective against neoplasia without producing substantial concomitant immunosuppression.
  • High dosages of cytochalasins may also be administered as sustained release formulations in microcapsules without producing immunosuppression.
  • a preferred type of microcapsule is that encapsulated in a film-forming polymer made of a mixture of polylactate-glycolate in a weight ratio of about 45:55 to about 55:45, preferably 50:50, and similar materials as taught by U.S. Patent Nos. 4,675,189, 4,585,482, 4,542,025, 4,530,840, European Patent Applications 87309286.0 (Publication Number 0,266,119), 87307115.3 (Publication Number 0257915), 81305426.9 (Publication Number 0052510) and 83303605.6 (Publication Number 0129619).
  • sustained release versions of cytochalasin in microcapsules may also be used in formulating sustained release versions of cytochalasin in microcapsules. Still other sustained release formulations readily recognized in the art may be used in this aspect of the present invention provided that the total amount of cytochalasin included within the formulation does not produce immunosuppression, i.e., is no greater than about three times the MTD of cytochalasin.
  • sustained release formulations containing cytochalasin may be used provided that the release of cytochalasin is such that otherwise immunosuppressive levels of cytochalasin are not reached rapidly at immunosuppressive sites, i.e., within a period of at least about 1 to 12 hours, and preferably within a period of at least about 1 to 24 hours.
  • Delivery systems other than liposomes or microcapsules may also be used for administering both natural and semi-synthetic cytochalasins as immunosuppressive or antineoplastic agents.
  • Such delivery systems include for example osmotic pumps, transdermal patches, infusion pumps, biodegradable polymers, monoclonal antibody-linked systems, suppositories, rhinile, dragees and troches, among others.
  • the present invention also relates to the use of IL-2 to eliminate the immunosuppression produced by cytochalasins. It has been discovered that the use of IL-2 will reverse the immunosuppressive effects of cytochalasins, providing an effective means of overcoming the immunosuppression that occurs when high doses of cytochalasins alone, or in combination with other antineoplastic agents, are administered for the treatment of neoplasia. In this aspect of the present invention, the administration of IL-2 to cytochalasin immunosuppressed lymphocytes will eliminate the immunosuppression.
  • IL-2 administered any time during immunosuppression will eliminate the immunosuppression, to maximize its effect, it is preferred that the IL-2 should be administered in conjunction with cytochalasin or within a short period of time thereafter, for example, within one hour.
  • the amount of IL-2 administered will vary depending upon the individual patient, the type of cytochalasin used and the extent of immunosuppression, preferred dosages of IL-2 range from about 5,000 to about 15,000 units per kilogram per day.
  • IL-2 from any source may be utilized in this aspect of the present invention, including IL-2 produced by cultivating human peripheral blood lymphocytes or other IL-2 producing cell lines, the preferred IL-2 is human recombinant IL-2 (rIL-2, available from DuPont Wilmington, Delaware).
  • any protein having IL-2 activity may be used to eliminate the immunosuppressive effects of the cytochalasins and that the term IL-2 includes proteins in which one or more of the amino acids of IL-2 have been changed but in which activity is substantially the same as IL-2.
  • human rIL-2 is.preferred.
  • the administration of IL-2 in appropriate concentrations may be used to restore immunity.
  • the administration of IL-2 may be used to regulate the immunosuppressive activity of administered cytochalasin. Such administration may occur during or after the administration of cytochalasin alone or in combination with other antineoplastic agents.
  • IL-2 may be made by cultivating human peripheral blood lymphocytes (PBL) or other IL-2 producing cell lines. IL-2 may also be made by recombinant DNA technology, which has afforded a means to produce muteins and other modified versions of naturally occurring IL-2 which may be used to practice the present invention.
  • PBL peripheral blood lymphocytes
  • IL-2 may also be made by recombinant DNA technology, which has afforded a means to produce muteins and other modified versions of naturally occurring IL-2 which may be used to practice the present invention.
  • a preferred IL-2, human rIL-2 may be purchased from a number of suppliers (Cetus Corp., Emeryville, Calif.).
  • IL-2 may be formulated with cytochalasin using any of the solvents, additives and methods described hereinabove, including liposomes and microcapsules for sustained release formulations, provided that the formulations do not disturb the integrity, stability or activity of the IL-2.
  • IL-2 may be administered in combination with cytochalasin or preferably, shortly thereafter, via parenteral routes of administration, preferably via IV infusion.
  • IL-2 and cytochalasin may also be administered in combination with at least one additional antineoplastic agent.
  • antineoplastic agents may be used in combination with cytochalasins and IL-2, including doxorubicin, daunorubicin or epirubicin, pyrrolizidine alkaloids, the vinca alkaloids such as vinblastine or vincristine, the purine or pyrimidine derivatives for example 5-fluorouracil, among others, the alkylating agents such as itoxanthrone, mechlorethamine hydrochloride or cyclophosphamide, platinum compounds such as cis-platinum, folic acid analogs such as methotrexate, and the antineoplastic antibiotics such as mitomycin or bleomycin, among others.
  • doxorubicin daunorubicin or epirubicin
  • pyrrolizidine alkaloids such as vinblastine or vincristine
  • the purine or pyrimidine derivatives for example 5-fluorouracil
  • the alkylating agents such as itoxanthrone,
  • the present invention also relates to a method and formulations for treating splenomegaly (enlarged spleen) in humans.
  • the same formulations of cytochalasin including the same amounts of cytochalasin used to induce immunosuppression are effective for treating splenomegaly resulting from a hyperimmune state.
  • Another aspect of the present invention relates to novel semi-synthetic analogues of CB which are not as readily metabolized to the enedione system of CA as is CB. Synthetic analogues of the present invention have the general structure
  • cytochalasin analogues of the present invention are commonly known as 20-deoxy-cytochalasin, 20-deo ⁇ y-21,21- dihydrocytochalasin, 20-deoxy-20-fluoro-cytochalasin and 20- deoxy-20-fluoro-21,22-dihydrocytochalasin.
  • the semi-synthetic cytochalasins of the present invention may not be as readily metabolized to the enedione system of CA, a system which is believed to be responsible for much of the toxicity associated with the administration of CB and CA.
  • the enedione system may irreversibly bind with any number of thiol groups and other nucleophiles in the biological system, thus producing toxic effects.
  • the semi-synthetic cytochalasins of the present invention are designed to maintain the activity associated with other cytochalasins such as CB, including immunosuppression, antineoplasia and anti-metastasis without readily metabolizing or converting, in vivo to the enedione system of CA.
  • the semi-synthetic cytochalasins of the present invention are advantageously longer-acting than CB or CA and are designed to reduce the marked deleterious side-effects associated with the administration of CB or CA.
  • the semi-synthetic analogues of the present invention may be made by a number of procedures including total chemical synthesis. However, the preferred route of synthesizing these analogues is to chemically modify CA which has been previously isolated from the mold D. dematioidea (ATCC 24346) or which has been prepared by the oxidation of CB to CA using standard prior art methods. Any of the known prior art methods for isolating CA or CB may be used, but it is preferred to utilize a batch absorption technique according to the procedure set forth in PCT Publication No.
  • CA is subjected to a blocking procedure in which the free hydoxyl group is blocked with, for example, a formyl group (85% HC00H at 60 C for one hour).
  • a formyl group 85% HC00H at 60 C for one hour.
  • the 20 keto group is subjected to a reduction to produce a hydroxyl at the C20 position.
  • the free hydroxyl at C20 is then activated with toluensulfonylchloride to produce the "tosyl" group at C20.
  • the free hydroxyl group of CA is blocked with the formyl group, and the resulting 0 product is subjected to a blocking procedure in which the 20-keto group is blocked as the ketal with, for example 1,3-dihydroxypropane in acid.
  • the activated double bond at the 21,22 position is then reduced in NaBH4 to produce the 21,22 dihydro derivative.
  • the 20 keto group may be reduced to form 21,22-dihydrocytochalasin B.
  • the free hydroxyl group may be first blocked with a formyl group and the 20 keto group reduced.
  • the resulting free hydroxyl group is o tosylated and the tosyl group displaced with iodine as described hereinabove to produce 20-iodo-21,22-dihydrocytochalasii_.
  • 20-iodo-21,22-dihydrocytochalasin may be converted to 20-deoxy-20-fluorocytochalasin by simple displacement of the iodo group with fluorine (KF or CsF/18-Crown 6 in acetonitrile).
  • the 5 20-fluoro cytochalasin derivative may be a diastereomeric mixture (racemic at C20), but the fluoro group at G20 is preferably of the same configuration as is the C20 hydroxyl of CB.
  • the hydroxyl group of CA is blocked with the formyl group, the 20 keto group reduced, tosylated and iodinated as described above.
  • the 20-iodocytochalasin is then converted to 20-deoxy-20-fluorocytochalsin by simple displacement of the iodo group at C20 with fluorine as described above.
  • the 20-fluoro cytochalasin mixture may be a diastereomeric mixture (the fluorine at €20 is a racemic mixture), but the fluoro group at C20 is preferably of the same configuration as is the C20 hydroxyl of CB.
  • the semi-synthetic cytochalasins of the present invention may be used as immunosuppressive agents or agents for treating neoplasia with or without IL-2 or additional antineoplastic agents.
  • the dosage that is to be administered to produce transient, reversible immunosupression is the same as for the other cytochalasins, i.e., at least about one half the MTD for a particular route of administration. Because of their expected longer duration activity, smaller amounts of semi-synthetic cytochalasins than CB are to be used to produced immunosuppression or to treat neoplasia.
  • the dosage of semi-synthetic cytochalasin for treating neoplasia preferably ranges from about one tenth the MTD to about one fifth the MTD when cytochalasin is to be administered alone or in combination with additional antineoplastic agents.
  • IL-2 may be administered in combination with the semi-synthetic cytochalasins.
  • the semi-synthetic cytochalasins may be administered via intravenous, intramuscular, sub-cutaneous and oral routes, according to the 0 general principles and methods and using the additives and solvents fully described hereinabove.
  • Sustained release formulations comprising very high dosages, e.g., within the range of about the MTD to about three times the MTD of the semi-synthetic cytochalasins of the present invention and liposomes or microcapsules are also 5 contemplated by the present invention.
  • Such dosages in sustained release form provide sufficient antineoplastic amounts of cytochalasin and produce surprisingly low amounts of immunosuppression.
  • the prescribing physician will ultimately determine the appropriate dosage of the cytochalasin and other agents including IL-2 or-neoplastic agents for a given human subject and condition, and this can be expected to vary according to the age, weight, and response of the individual as well as the nature and severity of the patient's disease or condition. Dosages would be ultimately determined by the administering physician according to the specific cytochalasin used, the circumstances of treatment and the pharmacokinetics of the agent in the patient.
  • the general dosage ranges provided herein should provide a guideline to follow in making the final dosage determinations.
  • the dosage of the drug in liposomal or microcapsule form for the treatment of neoplasia without immunosuppression will generally be up to about three times that employed for the free drug. In some cases, however, it may be necessary to administer dosages outside these limits.
  • Crystalline CB was subjected to final purification by preparative HPLC (75:25 MeOH/H20 on reverse phase C18 DYNAMAXtm silica) followed by recrystallization from CHC13. Purity was shown to be greater than 99% by analytical TLC and HPLC.
  • DBA/2 and C57BL/6 female mice were purchased from Charles River Laboratories, Wilmington, Massachusetts through the Animal Genetics Branch of the National Cancer Institute and used at 8 to 12 weeks of age.
  • CB was prepared for injection using emulsifying needles of decreasing bore diameter from 20 gauge to a final 25 gauge according to the method described in Bosquet, et al. submitted for publication. Briefly, suspensions were achieved by weighing CB into a syringe, loading the vehicle into a second syringe, and connecting the syringes with a double-hubbed emulsifying needle.
  • the suspension was forced back and forth repeatedly through the coupling tube until flow was unimpeded, then progressively the bore diameter was decreased from 21 gauge, to 23 gauge and finally to 25 gauge. Syringes and couplers were washed with MeOH and the MeOh wash analyzed on TLC to determine residual CB that was not actually delivered to the test animals.
  • the culture media for cell culture was RPMI 1640 medium with HEPES, fetal bovine serum, sodium pyruvate, gentamycin, penicillin/streptomycin, MEM non-essential amino acids and trypan blue, purchased from GIBCO, Grand Island, New York, USA.
  • RPMI 1640 medium with HEPES was supplemented for the 4-day sensitization cell cultures with 10% fetal bovine serum, 0.1 mg/ml gentamycin, 1% sodium pyruvate, 1% non-essential amino acids, 1% glutamine, and 50 uM mercaptoethanol. Medium was immediately sterilized by membrane filtration and used immediately or, if not used immediately, was directly sterilized prior to use.
  • RPMI 1640 medium used for the 4 hour Cr cytotoxicity assay was supplemented with 5% fetal bovine serum and 1% penicillin/streptomycin solution.
  • P815 mastocytoma tumor cells were maintained in ascites form by weekly intraperitoneal passage into syngeneic DBA/2 mice.
  • Tumor cells used for antigen in 4-day sensitization assays were harvested 72 hours after passage of 5 to 10 x 106 cells. The cells were washed, resuspended in RPMI 1640 medium with HEPES buffer and x-irradiated on ice in a 35 X 10 mm tissue culture dish at a rate of 200 rad/min and 10 cm target distance for a total of 21 minutes (4200 rad).
  • Cr51 labelled tumor used for targets in a 4 hour , cytotoxicity (LMC) assay were prepared according to the method of Cerottini, et al. (1972 Nature, 237, 272 and 1974 J. Exp. Med., 140, 703) with the modifications of Orsini, et al. (1977, Cancer Res., 37, 1719).
  • 3 X 107 P815 mastocytoma tumor cells were implanted IP into C57BL/6 mice previously treated or to be treated with IP CB at concentrations of 50 mg/kg or with vehicle (CMC/TW- control). The mice were injected with this dose at -24, -12. -6, -3, simultaneously with, or 24, 48 or 72 hours after implantation. 9 days after tumor challenge, splenectomy occurred.
  • spleens were aseptically ,5 removed from drug-treated and vehicle-treated (control) mice at 72 hours, 19 hours or three hours after injection with CB. Spleen cells were then forced through coarse (50 mesh) 'and fine (200 mesh) sterilized stainless steel wire mesh to make a single cell suspension. Cells were washed three times (except where indicated) 0 with RPMI 1640 medium and viability was determined by trypan blue exclusion.
  • the 4-day sensitization of splenic lymphocytes in the presence of x-irradiated P815 tumor antigen was carried out by the method of 5 Cerottini, et al. (1972 Nature, 237, 272 and 1974 J. Exp. Med., 140, 703) with the modifications of Orsini, et al. (1977, Cancer Res., 37, 1719).
  • Spleen cells were cultured in 17.8 X 35 mm 6- well tissue culture plates (Costar, Cambridge, Mass., USA) in a total volume of 2 ml. Each well contained 2 X 107 viable spleen cells and 0 4 X 105 x-irradiated P815 cells.
  • effector lymphocytes were recovered from each well by gentle scraping with a rubber policemen, and triplicate cultures were pooled.
  • Cells were washed with RPMI 1640 medium and counted with a hemacytometer or flow cytometer (Coulter Diagnostics, Hialeah, Fla., 5 USA).
  • the cytotoxic activity of sensitized spleen cells recovered from the 4 day cultrures was determined in a standard 4 hour 51Cr release assay as previously described by Bogyo and Mihich, Cancer Research, 40, 650 (1980). Each cell preparation was assayed with a minimum of four different effector:target cell ratios ranging from 100:1 to 6:1.
  • the percentage of 51Cr release was calculated according to the formula:
  • Human recombinant interleukin-2 (obtained as a gift from DuPont Corp, Wilmington, Del., USA) was added to unwashed cultured splenic cells from example 1 at a level of 10 U/ml in 100 microliters of RPMI 1640 medium at the time of culture initiation. Unwashed and washed spleen cells were used in order to determine whether restoration of immune-responsiveness by rIL-2 could be obtained under conditions where any endogenous CB, CB metabolites or suppressive factors remained present. At the end of the 4 day culture period effector lymphocytes were recovered from each well by scraping.
  • microencapsulated CB formed from a mixture of polylactate-glycolate under contract from Southern Research Institute- which may also be prepared according to the general method taught by U.S. Patent No. 4,389,330
  • Figure 10 shows that a dose of CB in microencapsulated form which is 3-fold above the maximum tolerated dose for IP bolus administration can be administered to mice without producing significant immunosuppression either 19 hours or 72 hours following injection.
  • CA is isolated from the mold D. dematioidea (ATCC 24346) or oxidized from CB isolated from that same mold according to the procedure set forth in PCT Publication No. WO 88/10259.
  • CA is subjected to blocking of the hydroxyl group with a formyl group by stirring CA in an excess of 85% HC00H at 60 C for one hour or acetylformate/pyridine at -20 C, isolated by ether extraction followed by evaporation of solvent.
  • the crude formyl blocked Cytochalasin A is then subjected to reduction in NaBH4/Dimethylformamide (DMF) or dimethylsulfoxide (DMSO) to produce the formyl blocked derivative having a free hydroxyl group at C20.
  • DMF NaBH4/Dimethylformamide
  • DMSO dimethylsulfoxide
  • the free hydroxyl group at C20 is tosylated (excess toluenesulfonylchloride/pyridine at 37 C overnight) and the tosylated product is extracted with ether after the excess pyridine is evaporated (several times with ethanol).
  • the tosylated product is then subjected to reaction with a 1.5 molar excess of Nal in DMF or DMSO to displace the tosyl group with an iodo group at C20, which is then subjected to reductive displacement of the iodo group with NaBH4 in DMF/DMS0 and deblocking of the formyl group under acidic conditions (6N HC1)) to produce 20-deoxycytochalasin.
  • the 20-iodocytochalasin derivative may be used to synthesize
  • 20-fluorocytochalasin in Example 9 20-fluorocytochalasin in Example 9. 20-deoxycytochalasin may be purified from the reaction mixture using preparative HPLC. It is sometimes preferable, but more time consuming, to isolate the intermediates of the synthesis using preparative HPLC (75:25 MeOH/H20 on reverse phase-C18 DYNAMAXtm silica gel columns) befpre proceeding on to the next synthetic step.
  • preparative HPLC 75:25 MeOH/H20 on reverse phase-C18 DYNAMAXtm silica gel columns
  • the free hydroxyl group of CA is blocked with the formyl group 85% HC00H at 60 C or acetylformate/pyridine at -20 C, and the resulting product is subjected to a blocking procedure in which the 20-keto group is blocked as the ketal with 1,3-dihydroxypropane or alternatively with 1,3-dimethoxypropane in toluenesulfonic acid.
  • the ketal-protected derivative is subjected to NaBH4 reduction (DMF or DMSO) or Pd/Charcoal/H2 reduction in ethanol to produce the ketal protected 21,22 dihydrocytochalasin.
  • the resulting product is then subjected to acid cleavage of the ketal (toluenesulfonic acid in dioxane) followed by blocking of the free hydroxyl group with a formyl group in 85% HC00H at 60 C for one hour or alternatively, acetylformate/pyridine at -20 C.
  • the 20-keto group is reduced to the hydroxy group, the hydroxy group is tosylated and the tosyl group is displaced by iodination (this iodo intermediate may be used to prepare the product of Example 11) reduced according to the method described in Example 8.
  • the formyl group is cleaved in 6N HCl.
  • the resulting product, 20-deoxy-21,22-dihydrocytochalasin may be purified from the reaction mixture using preparative HPLC (as described in example 8).
  • 20-deoxy-20-fluoro-21,22-dihydrocytochalasin may be produced from 20-iodo-21,22-dihydrocytochalsin from example 9 by simple displacement of the iodo group at C20 with KF or CsF/18 Crown 6 (1 molar equivalent of 18 Crown 6) in acetonitrile as per example 10 followed by removal of the formyl blocking group (6N HCl).
  • Preparative HPLC may be performed as previously described. It is , 0 to be understood that the examples and embodiments described hereinabove are for the purposes of providing a description of the present invention by way of example and are not to be viewed as limiting the present invention in any way. Various modifications or changes that may be made to that described hereinabove by those of 5 ordinary skill in the art are also contemplated by the present invention and are to be included within the spirit and purview of this application and the following claims.
  • the invention also contemplates new derivatives of cytochalasins as shown at page 22 wherein the formula for the oxidized or reduced
  • R , R., R. and R_ are hydrogen, hydroxy or halogen, wherein R and R_ or R. and R_ taken together are 0 oxygen, or wherein R- and R. when taken together are oxygen, and additionally wherein R. is hydrogen or fluoro.

Landscapes

  • Chemical & Material Sciences (AREA)
  • Health & Medical Sciences (AREA)
  • Organic Chemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • Veterinary Medicine (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Animal Behavior & Ethology (AREA)
  • Medicinal Chemistry (AREA)
  • Public Health (AREA)
  • Epidemiology (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • General Chemical & Material Sciences (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
  • Medicinal Preparation (AREA)
  • Preparation Of Compounds By Using Micro-Organisms (AREA)
  • Nitrogen Condensed Heterocyclic Rings (AREA)

Abstract

L'invention concerne des compositions de cytochalasine naturelles et synthétiques ainsi que des traitements thérapeutiques utilisant ces compositions. Plus particulièrement, l'invention concerne certains analogues synthétiques de cytochalasine B (CB) et des compositions à libération entretenue contenant une cytochalasine, par exemple la cytochalasine B et d'autres cytochalasines naturelles, par exemple la cytochalasine D, E, H ou J, entre autre, et un ou plusieurs de ces analogues synthétiques. L'invention concerne également la découverte surprenante, à savoir que l'administration de cytochalasines comprenant CD produit une immunosuppression transitoire qui est régulable par dose ou administration et est réversible spontanément ou en utilisant UL-2. Ainsi, un régime thérapeutique de cytochalasines peut être administré pour traiter l'hyperimmunité indésirable de patients ayant subis une transplantation et des patients atteints de maladies auto-immunes. De plus, une thérapie anti-tumeur utilisant CB et autres cytochalasines, et éventuellement des agents anti-néoplastiques autres que les cytochalasines peut être considérablement améliorée en combinant l'administration de ces agents avec des quantités efficaces de IL-2 ou autres lymphokines pour inverser l'immunosuppression produite pendant l'administration des cytochalasines ou des cytochalasines combinées à d'autres agents anti-néoplastiques. Cette invention concerne également des compositions à libération entretenue utilisant des liposomes ou des microcapsules qui sont efficaces pour apporter des concentrations élevées de cytochalasines et, éventuellement, des agents anti-néoplastiques supplémentaires au site actif de la tumeur sans produire une immunosuppression indésirable.
PCT/US1990/002342 1989-04-28 1990-04-27 Compositions de cytochalasine et procedes therapeutiques WO1990013293A1 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
US34501089A 1989-04-28 1989-04-28
US345,010 1989-04-28

Publications (1)

Publication Number Publication Date
WO1990013293A1 true WO1990013293A1 (fr) 1990-11-15

Family

ID=23353087

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/US1990/002342 WO1990013293A1 (fr) 1989-04-28 1990-04-27 Compositions de cytochalasine et procedes therapeutiques

Country Status (4)

Country Link
EP (1) EP0429585A4 (fr)
JP (1) JPH03505744A (fr)
CA (1) CA2031520A1 (fr)
WO (1) WO1990013293A1 (fr)

Cited By (17)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1993011763A1 (fr) * 1991-12-09 1993-06-24 Usher Thomas C Utilisation de cytochalasines pour inhiber la replication virale
US5250563A (en) * 1991-09-25 1993-10-05 Merck & Co., Inc. Inhibitors of HIV protease
US5342926A (en) * 1993-06-08 1994-08-30 The Regents Of The University Of California Analogs of cytochalasin B as radiopharmaceuticals for nuclear imaging of trans-membrane glucose transport
WO1999027908A1 (fr) * 1997-12-04 1999-06-10 Yissum Research Development Company Of The Hebrew University Of Jerusalem Traitement chimio et immunotherapeutique combine a l'aide de medicaments et de cytokines encapsules dans des liposomes
US6171609B1 (en) 1995-02-15 2001-01-09 Neorx Corporation Therapeutic inhibitor of vascular smooth muscle cells
US6268390B1 (en) * 1991-09-27 2001-07-31 Neorx Corporation Therapeutic inhibitor of vascular smooth muscle cells
US6787132B1 (en) * 1997-12-04 2004-09-07 Yissum Research Development Company Of The Hebrew University Of Jerusalem Combined chemo-immunotherapy with liposomal drugs and cytokines
US7094550B2 (en) 1993-05-13 2006-08-22 Neorx Corporation Method to determine TGF-beta
US7445628B2 (en) 1995-06-07 2008-11-04 Cook Incorporated Method of treating a patient with a coated implantable medical device
EP2098230A1 (fr) 1997-03-31 2009-09-09 Boston Scientific Scimed Limited Utlisation des inhibiteurs cytosquelettiques en forme cristalline pour l'inhibition ou la prévention de la resténose
EP0975340B2 (fr) † 1997-03-31 2009-10-28 Boston Scientific Limited Inhibiteur therapeutique des cellules des muscles lisses de la paroi vasculaire
US7625410B2 (en) 2001-05-02 2009-12-01 Boston Scientific Scimed, Inc. Stent device and method
US9066990B2 (en) 2001-03-26 2015-06-30 Bayer Intellectual Property Gmbh Preparation for restenosis prevention
US9649476B2 (en) 2002-09-20 2017-05-16 Bayer Intellectual Property Gmbh Medical device for dispersing medicaments
CN110563634A (zh) * 2019-09-09 2019-12-13 湖南省中医药研究院 吲哚细胞松弛素类化合物及其制备方法和应用
CN111249445A (zh) * 2020-02-27 2020-06-09 广东医科大学 一种细胞松弛素h软胶囊及其制备方法
US11007176B2 (en) 2019-02-19 2021-05-18 Tzu Chi University Use of actin depolymerizing agents for the treatment of anxiety disorders

Families Citing this family (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2009069668A1 (fr) * 2007-11-28 2009-06-04 National University Corporation Nagoya University Agent pour augmenter l'expression d'un antigène du mélanome malin, et son utilisation

Family Cites Families (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CA1289077C (fr) * 1984-08-13 1991-09-17 Harry H. Leveen Traitement du cancer a l'aide de la phlorizine et de ses derives
AU1969388A (en) * 1987-06-19 1989-01-19 Syracuse University Cytochalasin purification methods and compositions

Non-Patent Citations (2)

* Cited by examiner, † Cited by third party
Title
CHEMICAL ABSTRACTS, Volume 93, Number 25, issued 22 December 1980, E. PAUNESCU, "In Vivo and in Vitro Comparative Immunosuppressive Effect on T Cells of Cytochalasin B(cyt.B) Cyclophosphamide (CF), Actinomycin D (Act.D) and two Rifamycin (RF/SV) Derivatives." see page 44, column 1, Abstract number 230750U. *
See also references of EP0429585A4 *

Cited By (20)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5250563A (en) * 1991-09-25 1993-10-05 Merck & Co., Inc. Inhibitors of HIV protease
US6268390B1 (en) * 1991-09-27 2001-07-31 Neorx Corporation Therapeutic inhibitor of vascular smooth muscle cells
WO1993011763A1 (fr) * 1991-12-09 1993-06-24 Usher Thomas C Utilisation de cytochalasines pour inhiber la replication virale
US7094550B2 (en) 1993-05-13 2006-08-22 Neorx Corporation Method to determine TGF-beta
US5342926A (en) * 1993-06-08 1994-08-30 The Regents Of The University Of California Analogs of cytochalasin B as radiopharmaceuticals for nuclear imaging of trans-membrane glucose transport
WO1994028943A1 (fr) * 1993-06-08 1994-12-22 The Regents Of The University Of California Analogues de cytochalasine b utilises comme produits pharmaceutiques marques pour visualiser par imagerie nucleaire le transport du glucose a travers la membrane
US6171609B1 (en) 1995-02-15 2001-01-09 Neorx Corporation Therapeutic inhibitor of vascular smooth muscle cells
US7445628B2 (en) 1995-06-07 2008-11-04 Cook Incorporated Method of treating a patient with a coated implantable medical device
EP2098230A1 (fr) 1997-03-31 2009-09-09 Boston Scientific Scimed Limited Utlisation des inhibiteurs cytosquelettiques en forme cristalline pour l'inhibition ou la prévention de la resténose
EP0975340B2 (fr) † 1997-03-31 2009-10-28 Boston Scientific Limited Inhibiteur therapeutique des cellules des muscles lisses de la paroi vasculaire
EP2292225A1 (fr) 1997-03-31 2011-03-09 Boston Scientific Scimed Limited Forme de dosage comprénant du taxol en forme cristalline
US6787132B1 (en) * 1997-12-04 2004-09-07 Yissum Research Development Company Of The Hebrew University Of Jerusalem Combined chemo-immunotherapy with liposomal drugs and cytokines
WO1999027908A1 (fr) * 1997-12-04 1999-06-10 Yissum Research Development Company Of The Hebrew University Of Jerusalem Traitement chimio et immunotherapeutique combine a l'aide de medicaments et de cytokines encapsules dans des liposomes
US9066990B2 (en) 2001-03-26 2015-06-30 Bayer Intellectual Property Gmbh Preparation for restenosis prevention
US7625410B2 (en) 2001-05-02 2009-12-01 Boston Scientific Scimed, Inc. Stent device and method
US9649476B2 (en) 2002-09-20 2017-05-16 Bayer Intellectual Property Gmbh Medical device for dispersing medicaments
US11007176B2 (en) 2019-02-19 2021-05-18 Tzu Chi University Use of actin depolymerizing agents for the treatment of anxiety disorders
CN110563634A (zh) * 2019-09-09 2019-12-13 湖南省中医药研究院 吲哚细胞松弛素类化合物及其制备方法和应用
CN111249445A (zh) * 2020-02-27 2020-06-09 广东医科大学 一种细胞松弛素h软胶囊及其制备方法
CN111249445B (zh) * 2020-02-27 2022-11-15 广东医科大学 一种细胞松弛素h软胶囊及其制备方法

Also Published As

Publication number Publication date
JPH03505744A (ja) 1991-12-12
EP0429585A1 (fr) 1991-06-05
EP0429585A4 (en) 1992-03-25
CA2031520A1 (fr) 1990-10-29

Similar Documents

Publication Publication Date Title
WO1990013293A1 (fr) Compositions de cytochalasine et procedes therapeutiques
US4873088A (en) Liposome drug delivery method and composition
US5169934A (en) Intracellularly cleavable compounds
US7887836B2 (en) Compositions and methods for treating lymphoma
JP4524071B2 (ja) Ｓｎ−３８脂質複合体及び使用方法
JP3074733B2 (ja) 脂肪乳剤
CA2366787C (fr) Traitement du lymphome et composition a cet effet
HU214326B (hu) Eljárás felnőtt T-sejt leukémia/limfóma kezelésére alkalmas gyógyszerkészítmények előállítására
EP0036676A1 (fr) Procédé de préparation de liposomes de taille identique et les liposomes ainsi obtenus
JPWO2005117878A1 (ja) イリノテカン製剤
EP0297946A2 (fr) Méthodes de purification de cytochalasine et ses compositions
JP5295277B2 (ja) 配糖体含有リポソーム
US20020044951A1 (en) Isolated and purified nonpeptide antigens from mycobacterium tuberculosis
JP2003528908A (ja) Ｄおよびｌエーテル脂質立体異性体およびリポソーム
JP2010215662A (ja) １３−デオキシアントラサイクリン誘導体及びその製造方法
US6267968B1 (en) MDP derivatives and conjugates having haematipoietic function stimulating activity, and compositions containing same
KR0185226B1 (ko) 이식 조직의 거부반응 억제제 및 il-1 생산 억제제
JP2002535251A (ja) 癌治療のための新規化合物
KR20010101775A (ko) 글루타티온 유사체를 함유하는 치료 조성물
EP4268800A1 (fr) Liposome comprenant de la rapamycine ou un de ses dérivés et son utilisation en thérapie
US6121244A (en) Antitumoral method by administration of partricin derivatives
RU2121363C1 (ru) Иммуномодулятор и фармацевтическая композиция, обладающая противоопухолевой активностью
CA2255804C (fr) Methode antitumorale par administration de derives de partricine
JPH0516408B2 (fr)
WO2025037276A1 (fr) Compositions contenant des inhibiteurs de mtor et des lipides

Legal Events

Date Code Title Description
AK Designated states

Kind code of ref document: A1

Designated state(s): CA JP

AL Designated countries for regional patents

Kind code of ref document: A1

Designated state(s): AT BE CH DE DK ES FR GB IT LU NL SE

WWE Wipo information: entry into national phase

Ref document number: 2031520

Country of ref document: CA

WWE Wipo information: entry into national phase

Ref document number: 1990907829

Country of ref document: EP

WWP Wipo information: published in national office

Ref document number: 1990907829

Country of ref document: EP

WWW Wipo information: withdrawn in national office

Ref document number: 1990907829

Country of ref document: EP