HK1127026A - Bipolar trans carotenoid salts and their uses - Google Patents

Description

Bipolar trans carotenoid salts and uses thereof

The application is a divisional application, the original application of which has the application number of 03826969.4 and the application date of 2003, 8 and 25, and the invention is named as bipolar trans carotenoid salt and application thereof.

Technical Field

The present invention relates to bipolar trans carotenoid salt compounds, methods of dissolving, methods of making, and methods of using the same. These Bipolar Trans Carotenoid Salt (BTCS) compounds are useful for increasing oxygen diffusivity between red blood cells and body tissues in mammals including humans.

Background

Carotenoids are a class of hydrocarbons consisting of isoprenoid units linked in such a way that their arrangement is reversed in the center of the molecule. The main chain (backbone) of the molecule consists of conjugated carbon-carbon double and single bonds, and may also have pendant groups. Although the backbone of carotenoids has been thought to contain 40 carbons, for a long time it has been recognized that carotenoids may also have carbon backbones containing less than 40 carbon atoms. The 4 single bonds surrounding the carbon-carbon double bond all lie in the same plane. If the pendant group is on the same side of the carbon-carbon double bond, the group is said to be cis; if they are located on opposite sides of the carbon-carbon bond, they are said to be trans. Due to the large number of double bonds, carotenoids are highly likely to undergo geometric (cis/trans) isomerism, and are easily isomerised in solution. A recent series of books is the excellent bibliography for many of the properties of Carotenoids, etc. (compiled by Carotenoids ", G.Briton, S.Liaaen-Jensen and H.Pflander, Birkhauser Verlag, Basel, 1995, which is hereby incorporated by reference in its entirety).

Many carotenoids are non-polar and therefore insoluble in water. These compounds are very hydrophobic, which makes it difficult to formulate them for biological use, since organic solvents rather than aqueous solvents must be used in order to dissolve them. Other carotenoids are unipolar and have the characteristics of surfactants (hydrophobic moieties and hydrophilic polar groups). Thus, these compounds are attracted to the surface of the aqueous solution rather than being dissolved in the bulk liquid. There are small amounts of natural bipolar carotenoid compounds that contain a central hydrophobic portion and two polar groups, each at one end of the molecule. It has been reported ("Carotenoids", volume 1A, p 283) that carotenoid sulfates have significant solubility in water, reaching 0.4 mg/ml. Other carotenoids, which may be considered bipolar, are also not very soluble in water. These bipolar carotenoids include dialdehydes and diketones. Bipyridinium salts of crocetin have also been reported, but have a solubility in water of less than 1mg/ml at room temperature. Other examples of bipolar carotenoids are crocetin and crocin (both found in the spice crocus). However, crocetin is only slightly soluble in water. In fact, of all bipolar carotenes, only crocin shows significant solubility in water.

Us patent 4,176,179; 4,070,460, respectively; 4,046,880, respectively; 4,038,144, respectively; 4,009,270, respectively; 3,975,519, respectively; 3,965,261, respectively; 3,853,933 and 3,788,468 relate to various uses of crocetin.

U.S. Pat. No. 5,107,030 relates to a process for preparing 2, 7-dimethyl-2, 4, 6-octatrienedial and derivatives thereof.

U.S. Pat. No. 6,060,511 relates to Trans Sodium Crocetinate (TSC) and its use. The TSC is prepared by reacting naturally occurring saffron with sodium hydroxide followed by extraction.

In Roy et al, Shock 10, 213-217(1998), bleeding rats (55% blood volume) were given a bolus of Trans Sodium Crocetinate (TSC) 30 minutes later after the end of bleeding. All TSC treated animals survived, while all controls died. The systemic oxygen consumption increased in the TSC group to 75% of the normal resting value after about 15 minutes.

Laidig et al, J Am chem. Soc.120, 9394-9395(1998) relate to a computational model of TSC. The simulated TSC molecules were "hydrated" by surrounding the TSC molecules with water molecules. The rearrangement of water near the TSC allows oxygen molecules to diffuse more easily through the system. The calculated diffusivity increased by about 30%, which is consistent with the results obtained in vitro as well as in animal experiments.

In Singer et al, Crit Care Med 28, 1968-. Hypoxia was induced with an air mixture of low oxygen concentration (10%): animals were given saline or TSC after 10 minutes. Hypoxia results in reduced blood flow and increased base loss. Only 2 of the 6 animals of the control group survived. The treated group survived and had good hemodynamic stability for more than two hours, followed by a slow decline.

Disclosure of Invention

The present invention relates to Bipolar Trans Carotenoid Salt (BTCS) compounds and the synthesis of such compounds, said compounds having the following structure:

YZ-TCRO-ZY

wherein:

y is a cation;

z ═ a polar group bound to the cation, and

TCRO ═ trans carotenoid skeleton.

The present invention also relates to compositions of individual BTCS compounds (including compositions of TSC) wherein the absorbance of the highest peak of an aqueous solution of the BTCS composition occurring in the visible wavelength range divided by the absorbance of the peak occurring in the ultraviolet wavelength range is greater than 7.0, preferably greater than 7.5, most preferably greater than 8.

The present invention also relates to a method of treating a variety of diseases comprising administering to a mammal in need of treatment a therapeutically effective amount of a compound having the formula:

YZ-TCRO-ZY。

the present invention also includes dissolving and synthesizing a compound having the formula: several methods of preparing compounds of YZ-TCRO-ZY.

The invention also relates to an inhaler for delivering the compounds of the invention.

Detailed Description

A new class of carotenoids and carotenoid-related compounds has been discovered. These compounds are known as "bipolar trans carotenoid salts" (BTCS).

Compounds of the invention

The present invention relates to a class of compounds, namely bipolar trans carotenoid salts, which allow hydrophobic carotenoids or carotenoid-related skeletons to dissolve in aqueous solutions; and methods for their preparation. The cation of these salts can be of many kinds, but sodium or potassium (which are found in most biological systems) are preferred. Commonly owned U.S. patent 6,060,511, which is incorporated herein by reference in its entirety, describes an extraction process for the preparation of Trans Sodium Crocetinate (TSC), a BTCS, starting from saffron.

The general structure of bipolar trans carotenoid salts is:

YZ-TCRO-ZY

wherein:

y (which may be the same or different at both ends) ═ cation, preferably Na⁺、K⁺Or Li⁺. Y is preferably a monovalent metal ion. Y may also be an organic cation, e.g. R₄N⁺、R₃S⁺Wherein R is H or C_nH_2n+1Wherein n is 1 to 10, preferably 1 to 6. For example, R may be methylEthyl, propyl or butyl.

Z (which may be the same or different at both ends) is a polar group bound to the cation. Optionally including a terminal carbon on the carotenoid (or carotenoid related compound), which group may be a carboxyl group (COO)^-) Or a CO group. The radical may also be a sulfate radical (OSO)₃ ^-) Or a monophosphate group (OPO)₃ ^-)、(OP(OH)O₂ ^-) A diphosphate group, a triphosphate group, or a combination thereof.

TCRO ═ a linear trans carotenoid or carotenoid-associated backbone (preferably less than 100 carbons) having pendant groups (defined below) and typically comprising "conjugated" or alternating carbon-carbon double and single bonds (in one embodiment, TCRO is not fully conjugated as in lycopene). The pendant groups are typically methyl groups, but may be other groups as discussed below. In a preferred embodiment, the units of the backbone are linked in such a way that their arrangement is reversed in the center of the molecule. The 4 single bonds surrounding the carbon-carbon double bond all lie in the same plane. If the pendant group is on the same side of the carbon-carbon double bond, it is said to be cis; if the pendant group is located on the opposite side of the carbon-carbon double bond, it is referred to as trans. The compounds of the invention are trans. The cis isomer is generally detrimental and does not contribute to diffusivity. In one embodiment, trans isomers may be used whose backbone remains linear.

Examples of trans carotenoids or carotenoids-related skeletons are:

wherein the pendant groups X (which may be the same or different) are hydrogen atoms (H); or a linear or branched group having 10 or less than 10 carbons (preferably 4 carbons or less than 4 carbons) optionally containing a halogen; or a halogen. Essence of XFor example, methyl (CH)₃) (ii) a Ethyl (C)₂H₅) (ii) a Halogen-containing alkyl (C1-C10) such as CH₂Cl; or a halogen such as Cl or Br. The pendant groups X may be the same or different, but the X groups used must maintain a linear backbone.

Although many carotenoids exist in nature, no carotenoid salts exist. Commonly owned U.S. Pat. No. 6,060,511 relates to Trans Sodium Crocetinate (TSC). The TSC is prepared by reacting naturally occurring saffron with sodium hydroxide and then by extraction, which selects primarily the trans isomer.

The presence of cis and trans isomers of BTCS can be determined by observing the uv-vis spectrum of a carotenoid sample dissolved in an aqueous solution. Given a spectrum, the value of the absorbance of the highest peak occurring in the visible wavelength range of 416nm to 423nm (the number depending on the solvent used) divided by the absorbance of the peak occurring in the ultraviolet wavelength range of 250nm to 256nm can be used to determine the purity level of the trans isomer. When BTCS is dissolved in water, the highest peak in the visible wavelength range will be located at about 421nm and the peak in the UV wavelength range will be located at about 254 nm. The result (in that case the crocetin was analyzed) was calculated to be 3.1 according to m.craw and c.lambert, Photochemistry and Photobiology, volume 38(2), 241-243(1983), which is hereby incorporated by reference in its entirety, and increased to 6.6 after purification.

The Craw and Lambert analysis was performed on the trans sodium crocetinate of commonly owned U.S. patent 6,060,511 (TSC prepared by reacting naturally occurring crocetin with sodium hydroxide followed by extraction with a major selection of the trans isomer) using a sample cell designed for the UV and visible wavelength ranges, with the resulting value averaging about 6.8. This test is performed on synthetic TSCs of the present invention with the ratio being greater than 7.0 (e.g., 7.0 to 8.5), preferably greater than 7.5 (e.g., 7.5 to 8.5), and most preferably greater than 8. For TSC synthesized according to the modified method of example 5, this ratio is greater than 7.4 (e.g., 7.4 to 8.5). The material synthesized is a "more pure" or highly purified trans isomer.

It has recently been found that TSC has a water solubility of about 10mg/ml at room temperature, which is of great concern for molecules containing such long hydrophobic moieties. TSC has also been found to increase the diffusivity of oxygen through liquids.

U.S. Pat. No. 6,060,511 describes an extraction method for preparing TSC starting from saffron; however, other bipolar carotenoid salts cannot be prepared using the same procedure, since the use of saffron allows only a single carotenoid skeleton to be inserted into the salt.

The invention disclosed herein allows the synthesis of a whole class of compounds: a bipolar trans carotenoid salt containing a plurality of carotenoids and a carotenoid-related skeleton. Such compounds are soluble in aqueous solutions and have advantageous biological uses, for example leading to increased oxygen utilization. This increase is believed to be due to the ability of the hydrophobic portion (backbone) of the bipolar trans carotenoid to influence the binding of water molecules. This effectively causes the oxygen molecules to diffuse more rapidly in that region.

Dissolving the Compounds and compositions of the invention

The present invention allows trans carotenoids or carotenoid-related backbone molecules to be dissolved in aqueous solutions. The new dissolution method will be described below. The method is applicable to any bipolar trans carotenoid salt and compositions thereof.

Saline perfusion fluid containing BTCS

Large amounts (3 times the estimated blood loss) of isotonic saline (also known as normal saline) are infused as a treatment for hemorrhagic shock. The isotonic saline contains 9 grams NaCl per liter of water so that it does not destroy the ionic strength of the plasma once infused into the body. The addition of TSC to this saline has been shown to produce a better infusion solution, but such a solution cannot be prepared simply by mixing TSC powder with the saline. Regardless of how much TSC is added, about 50% of the TSC dissolves in physiological saline (up to a few milligrams per milliliter), which means that undissolved TSC particles are still present. To prevent this, a mother liquor can be prepared by adding more than twice the amount of TSC needed and then centrifuging out undissolved particles. The actual composition of the mother liquor can be identified using UV-visible spectroscopy. The mother liquor may be added to physiological saline and the TSC remains dissolved.

This method can be used to dissolve BTCS in other types of sodium chloride solutions and solutions such as KCl, Na₂SO₄And lactate, and other salts. Small amounts, e.g. 1mg/ml to 3mg/ml, can be added to the solution in this way.

Dissolving BTCS by dilute sodium carbonate solution

BTCS, such as TSC, is dissolved in an extremely dilute sodium carbonate solution. A dilute solution of, for example, 0.00001M to 0.001M sodium carbonate may be added dropwise to deionized water to a pH of 8.0 (the pH of deionized water is typically 5 to 6). For example, 50ml of deionized water requires only a few drops of extremely dilute sodium carbonate. This sodium carbonate-deionized water solution was able to completely dissolve large amounts of TSC (about 10mg/ml), which is of interest in view of the hydrophobicity of the carotenoid portion of BTCS.

BTCS can be provided as a powder with sterile bottled aqueous sodium carbonate solution. This concentrate can then be injected directly (very small amounts of a solution with a lower ionic strength than plasma can be injected), or the concentrate can be added to physiological saline and then injected. The TSC remains in solution if the TSC is dissolved in a sodium carbonate-water solvent and then more of the same solvent is added.

In another embodiment, sodium bicarbonate is used instead of sodium carbonate. Salts that result in deionized water having an alkaline pH may also be used.

A carotenoid skeleton concentration of 5mg/ml to 10mg/ml can be obtained by this procedure.

Water dissolution of BTCS

Although TSC was soluble in water (tap water, etc.),Distilled, deionized water), these solutions are stable only when the pH is adjusted so that the solution is basic. TSC in deionized water (with very little Na present)⁺Ions) are more soluble than in normal water. BTCS, such as TSC, is only dissolved in moderately deionized water and if pure deionized water is added to the solution, the TSC will precipitate out. BTCS dissolves only in moderately deionized water, but if the pH is not adjusted to be slightly alkaline, additional deionized water can cause precipitation of BTCS.

Other methods of dissolving BTSC

The BTCS may be formulated in a delivery system that enhances delivery. See below for the formulation of the compounds of the invention.

Synthesis of bipolar trans carotenoid salts of the invention

Described below are novel synthetic methods that can be used to synthesize bipolar trans carotenoid salts. Variations in the individual steps of the synthesis may be apparent to those skilled in the art.

TSC Synthesis

Can be prepared by reacting a symmetrical C containing conjugated carbon-carbon double bonds₁₀Dialdehydes (2, 7-dimethylocta-2, 4, 6-triene-1, 8-dialdehyde) and [ 3-carbomethoxy-2-buten-1-ylidene]The trans-sodium crocetinate (TSC) is synthesized by phase coupling of triphenylphosphine. This results in the formation of the trans dimethyl ester of crocetin. The dimethyl ester is then converted to the final TSC product by saponification. Typically, saponification is accomplished by treating the ester with aqueous sodium hydroxide or sodium hydroxide dissolved in THF (tetrahydrofuran); however, these methods do not produce optimal results in this case. In this case, saponification can be accomplished very well by reacting the ester with NaOH/methanol solution. After saponification, the TSC was recovered by vacuum drying.

C used in this synthesis₁₀The dialdehyde and triphenylphosphine reactants can be prepared via different routes. For example, from bromoacetic acid using Wittig chemistryStarting with ethyl ester and furan to prepare said C₁₀A dialdehyde. Tiglic acid is the starting material for the preparation of the desired phosphorane. By mixing different lengths of reactants (e.g. C)₁₄Dialdehyde and triphenylphosphine) were linked together to prepare carotenoid backbones of different lengths. This procedure results in the formation of different trans bipolar carotenoid salts. Variations can also be made to obtain different pendant groups (the pendant groups of the TSC have methyl groups).

TSC prepared in this way was dissolved in water at room temperature at a level of > 10mg/ml (pH adjusted to 8.0 with extreme dilution of sodium carbonate). Other bipolar trans carotenoid salts are soluble in water having a neutral or higher pH at room temperature. As used herein, "soluble" means an amount that is more than 5mg dissolved in water per ml at room temperature (as mentioned earlier, the carotenoid references indicate that 0.4mg/ml is "very significant solubility", but this is lower than the solubility defined in the present invention).

B. General Synthesis

Carotenoids or carotenoid-related structures can be constructed in the following manner:

(3-carbomethoxy-2-buten-1-ylidene) triphenylphosphorane (or related compound when X is not methyl) is a key precursor for adding isoprenoid units (or isoprenoid-related units) to both ends of a symmetrical carotenoid (or carotenoid-related compound). This process can be repeated indefinitely. For example, trans dimethyl crocylate can be reduced to the corresponding symmetrical dialdehyde using the chemistry described above. The dialdehyde can be reacted with an excess of (3-carbomethoxy-2-buten-1-ylidene) triphenylphosphorane to provide the corresponding diester. This synthesis sequence may be repeated iteratively.

Improved synthesis

2, 7-dimethyl-2, 4, 6-octatrienedial (2, 7-dimethylocta-2, 4, 6-triene-1, 8-dialdehyde) was used as a key intermediate for the synthesis of TSC. This key precursor has 3 double bonds, so several isomers may exist. For TSC, the all-trans isomer (E, E-isomer) is necessary. The conventional synthetic route involves 11 steps, the yield is relatively low, and the selectivity is low in several steps (see example 1). As a result, column chromatography must be used concomitantly with this route to purify several intermediates.

The improved synthetic route is much simpler (see reaction scheme below). The three-step process described in U.S. patent 5,107,030 (incorporated herein by reference in its entirety) provides a mixture of geometric isomers of the dialdehyde (this mixture is not noted in U.S. patent 5,107,030). In the process of the invention described in example 1, 96% to 97% of the desired isomer (all-trans or E, E, E-isomer) is obtained in 59% yield from methanol or ethyl acetate by several recrystallizations.

The improved synthesis of the present invention comprises the use of sulfinic acid (RSO) in a suitable solvent₂H, where R is a linear or branched alkyl group of C1 to C10 or an aryl group (substituted phenyl)) such as p-toluenesulfinic acid, the residual isomeric mixture of dialdehydes is converted by isomerization to the desired trans aldehyde (E, E), such as 1, 4-dioxane, tetrahydrofuran or dialkyl ethers in which the alkyl group is one or two of the linear or branched alkyl groups of C1 to C10. An additional 8% yield of pure desired dialdehyde was obtained, increasing the overall yield of the last step from 59% to 67%. This increase in yield is important. This isomerization step can be added to the third step of the process of U.S. patent 5,107,030 to obtain higher yields.

Improved synthetic route:

two undesired isomers:

isomerization of the undesired dialdehyde to the desired dialdehyde:

the diester can be prepared by dissolving the diester in methanol and then adding a base such as NaOH (in this case Y for BTCS is Na)⁺) To complete saponification. Alternatively, the diester may be dissolved in methanol already containing the base. Typically, NaOH is an aqueous solution (20 wt% to 60 wt%), but can also be a solid. Alternative to methanol for dissolving the diester are ethanol, propanol and isopropanol. Saponification can be accomplished in a variety of commercial ways. One or two phase systems (an organic phase and an aqueous phase) may be used.

Trans crocetin can also be synthesized according to the methods described above.

Furthermore, as has been reported for TSC, these BTCS compounds increase the diffusivity of oxygen through water (which will also depend on the nature of the hydrophobic moiety inserted into the end product, e.g. carbon chain length), since the hydrophobic interaction of the carotenoid skeleton with water is believed to result in an increase in diffusivity.

Preparation of the Compounds of the invention

A concentrate of bipolar trans carotenoid salts can be prepared as described above by dissolving it in an extreme dilution of sodium carbonate. The resulting mixture can then be used in that manner, or can be further diluted with physiological saline or other aqueous solvents. Furthermore, the solution of the bipolar carotenoid salt may be prepared by dissolving the bipolar carotenoid salt directly in a salt solution and subsequently removing any insoluble matter.

The dried forms of the bipolar trans carotenoid salts are stable at room temperature and can be stored for long periods of time. Preferably, the formulation of this salt, if an oral formulation, is absorbed in the intestinal tract rather than in the stomach.

Although the compounds of the present invention may be administered alone, they may also be administered as part of a pharmaceutical formulation. Such formulations may include pharmaceutically acceptable carriers as well as other therapeutic agents known to those skilled in the art (see below). Preferably, the formulation does not include a compound that inhibits the ability of the compound of the present invention to increase the diffusion of oxygen.

The appropriate dosage of the compounds and compositions of the present invention will depend on the severity of the condition being treated. For a "therapeutically effective" dose, it must have the desired effect, i.e., increase the diffusivity of oxygen. This will in turn cause the oxygen related parameter to return towards normal values.

Administration may be by any suitable route, including oral, nasal, topical, parenteral (including subcutaneous, intramuscular, intravenous, intradermal and intraosseous), vaginal or rectal. The preferred route of administration will depend on the details. Treatment by the inhalation route is preferred in emergency situations where the BTCS must enter the bloodstream very rapidly. The formulations thus include those suitable for administration via such routes (liquids or powders to be aerosolized). It will be appreciated that the preferred route may vary, for example, with the condition and age of the patient. The formulations may conveniently be presented in unit dosage form, for example, as tablets and sustained release capsules, and may be prepared and administered by methods well known in the art of pharmacy. The formulations may be used for immediate or sustained or controlled release of BTCS. See, for example, the controlled release formulation of WO 99/15150 (which is incorporated herein by reference in its entirety).

Formulations of the present invention suitable for oral administration may be presented as discrete units such as pills, capsules, cachets or tablets, as powders or granules, or as solutions, suspensions or emulsions. Formulations suitable for oral administration further include lozenges, pastilles and aerosols for inhalation in a suitable base or liquid carrier. Formulations for topical application to the skin may be in the form of ointments, creams, gels, and pastes containing the active agent and a pharmaceutically acceptable carrier, or in a transdermal patch.

Formulations suitable for intranasal administration in which the carrier is a solid include powders of a particular size which can be administered by rapid inhalation through the nasal passage. Suitable formulations in which the carrier is a liquid may be administered, for example as an intranasal spray or as drops.

Formulations suitable for parenteral administration include aqueous or non-aqueous sterile injection solutions which may contain anti-oxidants, buffers, bacteriostats and solutes which render the formulation isotonic with the blood of the intended recipient, and aqueous or non-aqueous sterile suspensions which may include suspending agents and thickening agents. The formulations may be presented in unit-dose or multi-dose containers, for example sealed ampoules and vials; and may be lyophilized, requiring only the addition of a sterile liquid carrier, e.g., water for injection, immediately prior to administration. Injection solutions and suspensions can be prepared from sterile powders, granules, and tablets.

Use of the Compounds and compositions of the invention

A wide variety of conditions are controlled or mediated by the delivery of oxygen to body tissues. The compounds and compositions of the invention can be used in the same pharmaceutical applications as those of crocetin in the same effective amounts; see U.S. patent 4,176,179; 4,070,460, respectively; 4,046,880, respectively; 4,038,144, respectively; 4,009,270, respectively; 3,975,519, respectively; 3,965,261, respectively; 3,853,933 and 3,788,468; these documents are incorporated herein in their entirety by reference.

TSC has been shown to increase the diffusivity of oxygen through aqueous solutions by about 30%. TSC increases survival in mammals following hypoxia, increases oxygen consumption following hypoxia or physiological stress, increases blood pressure following hypoxia, reduces post-hypoxia blood acidosis (i.e., reduces blood base deficiency, increases blood pH, and lowers plasma lactate levels), and reduces organ damage following hypoxia (e.g., liver, kidney damage). Thus, the compounds of the present invention are useful for treating diseases/disorders characterized by hypoxia (hypoxia) in mammals (including humans), such as, among others, respiratory diseases, hemorrhagic shock and cardiovascular diseases, multiple organ failure (e.g., due to ARDS sepsis or hemorrhagic shock), chronic renal failure, atherosclerosis, emphysema, asthma, hypertension, cerebral edema, papilloma, spinal cord injury, stroke, and the like. The compounds of the present invention are also useful for treating mammals at risk of developing the above-mentioned diseases/conditions. Other bipolar trans carotenoid salts have similar properties. Such compounds may also be used in combination with other methods commonly suggested for increasing oxygen utilization in the body, such as oxygen therapy and hemoglobin or fluorocarbons.

In one embodiment of the invention, the BTCS is administered to the patient at the same time as the oxygen is administered. Alternatively, hemoglobin or fluorocarbon, and BTSC may be administered together. In these cases, an additive effect is achieved.

The minimum dose of any of these salts required for treatment is the dose at which the diffusivity of oxygen is increased. The effective dosage of the compounds of the present invention will depend upon the condition being treated, the severity of the condition, the stage and individual characteristics exhibited by each mammalian patient. However, the dosage will vary from about 0.001mg to about 500mg of active compound per kg of body weight, preferably from about 0.01mg to about 30mg per kg of body weight. Intravenous administration is preferred, but other routes of injection such as intramuscular, subcutaneous or via inhalation may also be used. Transdermal or intraosseous delivery may be used, and oral administration may also be used.

Respiratory diseases

Bipolar trans carotenoid salts are also useful in the treatment of acute and chronic respiratory disorders. These are described as conditions in which the arterial oxygen partial pressure is reduced, for example, at values of 60mm Hg to 70mm Hg rather than the normal values of 90mm Hg to 100mm Hg. Such acute and chronic respiratory disorders include emphysema, polar lung injury (ALI), polar respiratory distress syndrome (ARDS), Chronic Obstructive Pulmonary Disease (COPD) and asthma.

When the partial pressure of oxygen in the blood is low (which is a symptom of emphysema, ARDS, and COPD), the TSC increases the value of the partial pressure of oxygen in the blood. Increasing the partial pressure of oxygen in the blood reduces many of the symptoms of emphysema, ARDS and COPD. TSC does not treat the etiology but alleviates oxidative stress and injury due to the underlying etiology.

Hemorrhagic shock

Hemorrhagic shock is marked by a decrease in oxygen consumption. Bipolar trans carotenoid salts increase the oxygen consumption of the body by allowing more oxygen to diffuse from the red blood cells into the tissue. TSC has been shown to increase oxygen consumption in rats in hemorrhagic shock, but it has also been shown to counteract other symptoms of shock. The compounds of the present invention cause elevated hypotension, a reduced accelerated heart rate and a reversal of the blood acidotic condition that develops during shock. The compounds of the invention also reduce organ damage following hemorrhagic shock.

The compounds of the invention may be administered by inhalation, injection or addition to standard resuscitation solutions (ringer's lactate or saline) for the treatment of hemorrhagic shock.

Cardiovascular diseases

In western civilization, the leading cause of death is ischemic heart disease. Death may be caused by a gradual decline in the ability of the heart to contract or often by sudden cessation of the heart. Sudden Cardiac Death (SCD) includes a period of time from 60 seconds after onset of symptoms to the following 24 hours. These deaths are often the result of acute coronary artery occlusion (obstruction) or ventricular fibrillation (which may result from occlusion).

Myocardial ischemia exists when insufficient oxygen is supplied to the myocardium. When coronary blood flow is very low, the myocardium is unable to function and dies. This muscle area is called infarct. Most often, the reduction in coronary blood flow is due to atherosclerosis occurring in the coronary arteries. Ischemia results in impaired mechanical and electrical performance and muscle cell damage, which can lead to fatal arrhythmia, known as Ventricular Fibrillation (VF). In ventricular fibrillation, the electrical activity of the heart ventricles is disordered and the rhythm of the resulting electrocardiogram is irregular and without recognizable patterns. Ventricular fibrillation is often accompanied by myocardial ischemia and infarction, and is almost always the cause of sudden cardiac death. Bipolar trans carotenoid salts are advantageous in the treatment of myocardial ischemia. Atherosclerosis and congestive heart failure, which are often precursors to myocardial infarction, may also be treated with these salts.

Ischemia of blood

Bipolar trans carotenoid salts are also advantageous in the treatment of other forms of ischemia (insufficient blood flow to tissues or organs), such as renal, hepatic, spinal and cerebral ischemia (including shock).

Surgery

Surgery often involves blood loss or artery clamping (e.g., bypass surgery), which may cause ischemia. Bipolar trans carotenoid salts are advantageous as a pretreatment for surgery or as an intra-or post-surgical treatment.

Hypertension (hypertension)

Hypertension or hypertension is often associated with cardiovascular disease. The compounds of the present invention are useful for lowering blood pressure.

Functional enhancement

BTCS enhances aerobic metabolism, increases oxygen consumption during walking, running, weight lifting, etc., and also increases tolerance.

Traumatic brain injury

Ischemia following traumatic brain injury exacerbates the brain injury. BTCS increases oxygen levels in brain tissue following impact injury (localized or diffuse injury). Examples of impact injuries include car/motorcycle accidents and falls. BTCS also increases the amount of oxygen reaching normal brain tissue when hyperoxic therapy is used.

Alzheimer's disease

In Alzheimer's disease, BTCS increases the level of oxygen consumption in the brain, thus alleviating the symptoms of Alzheimer's disease. Blood flow and oxygen consumption dropped to less than about 30% of the levels observed in non-demented elderly, Wurtman, Scientific American, volume 252, 1985.

The increased levels of oxygen consumption in the brain caused by BTCS also reduces memory loss.

Diabetes mellitus

BTCS is beneficial for the treatment of diabetic complications such as ulcers, gangrene and diabetic retinopathy. Diabetic foot ulcers, M.Kalani et al, Journal of Diabetes & Its formulations, Vol 16, No.2, 153-.

BTCS also contributes to complications of diabetic retinopathy associated with hypoxic stress, Denninghoff et al, Diabetes Technology & Therapeutics, Vol.2, No.1, 111-113, 2000.

Other uses

Bipolar trans carotenoid salts can also be used to treat spinal cord injury, cerebral edema, anemia and cutaneous papillomas. In all cases, they alleviated the disease, which was alleviated. This is believed to be a result of the increased oxygen consumption resulting from the use of bipolar trans carotenoid salts.

In addition, bipolar trans carotenoid salts can also be used to augment other physiologically important molecules such as glucose, CO₂Or diffusion of NO. BTCS alsoScavenging free radicals derived from oxygen.

*****

The following examples are illustrative, but not limiting, of the compositions and methods of the present invention. Other suitable modifications and adaptations of the numerous conditions and parameters which occur generally to those skilled in the art are within the spirit and scope of the invention.

Examples

Example 1

Synthesis of trans-sodium crocetinate

By combining a symmetrical C form containing conjugated carbon-carbon double bonds₁₀Dialdehydes and [ 3-carbomethoxy-2-buten-1-ylidene]The trans-sodium crocetinate is synthesized by coupling the triphenylphosphine phase. This product was then saponified with a NaOH/methanol solution.

Triphenylphosphine (at a concentration of about 2mol/L) dissolved in ethyl acetate was slowly added to ethyl bromoacetate. After separation and treatment with a base, the product may be treated with methyl iodide followed by treatment with a base to form the phosphorane. In this case, the basic compound to be formed into the carotenoid skeleton may be prepared starting from a cyclic compound such as furan. Furan is reacted with bromine and methanol, followed by a selective deprotonation step to form the monoaldehyde. The monoaldehyde is then coupled with a phosphorane. Acidic conditions deprotect other dimethylacetal groups to provide a free aldehyde. This compound is then reacted again with the same phosphorane described above to produce the diethyl diester. The ester group is reduced to an alcohol and subsequently oxidized (e.g. with MnO)₂Oxidation) to give C in the form of a dialdehyde₁₀And (3) a framework. The dialdehyde was subsequently reacted with a phosphorane prepared from tiglic acid. Esterifying the tiglic acid with methanol under acidic conditions to produce a methyl esterFollowed by a bromination step. The allylic bromide isomer is formed and can be isolated by crystallization. The desired bromide is then treated with sodium hydroxide to provide the desired phosphorane. Then the phosphorane and said C₁₀The dialdehyde is dissolved in a solvent such as toluene or benzene and refluxed. After removal of the solvent, the resulting product was isolated as a powder and subsequently saponified with a 40% NaOH/methanol mixture to form TSC.

***

Trans sodium crocetinate 1(TSC) was prepared in a 17-step synthetic sequence with an overall yield of 1.5%. A total of 4.1g TSC was prepared using ethyl bromoacetate, furan and tiglic acid as starting materials.

Trans Sodium Crocetinate (TSC) was synthesized from the saponification of dimethyl crocetinate, which was prepared based on the saponification of Buchta and Andree¹Total synthesis reported. The synthesis strategy after preparation of crocetin dimethyl ester is based on symmetric C₁₀Coupling of dialdehyde (2, 7-dimethylocta-2, 4, 6-triene-1, 8-dialdehyde) with (3-carbomethoxy-2-buten-1-ylidene) triphenylphosphine.

Although the original Buchta and Andree articles¹The title of (1) is "The Total Synthesis software range-2, 2-Bisdimethyl-Crocetin-dimethyl ester and trans-Crocetin-dimethyl ester", but The article does not report The details of The test and The yield. Found to be suitable for generating C after extensive search of documents₁₀Procedure for each step of dialdehyde and phosphorane. Finally, ethyl bromoacetate, furan and tiglic acid are used as starting materials according to 17 stepsThe sequence of (a) to make TSC with an overall yield of 1.5%.

Ethyl bromoacetate from Wittig chemical process²And furan³Preparation of said C₁₀A symmetrical dialdehyde. Treatment of ethyl bromoacetate with triphenylphosphine and iodomethane afforded the phosphorane 6:

a.TPP、EtOAc，92％；b.1 N NaOH、CH₂Cl₂；c.CH₃I、CH₂Cl₂；d.1 NNaOH、CH₂Cl₂。

the yield of the first step was quite high, 92%. The quantification of the subsequent steps in this sequence is complicated by the nature of the phosphorane 4 and the phosphonium salt 5. Both of these compounds are very viscous slurries, which produce a large amount of foam when concentrated in a rotary evaporator. Both compounds can be conveniently treated as dichloromethane solutions, and the overall yield of phosphorane 6 is qualitatively acceptable (estimated to be over 75%).

Furan is ring opened with bromine to provide the fumaryl bis (dimethyl acetal) 8.³

e.Br₂、MeOH、Na₂CO₃77%; f. macroporous resin (Amberlyst)15, H₂O, acetone, 72 percent.

Monodeprotection of bis (dimethylacetal) 8 under acidic conditions⁴Aldehyde 9 is generated and then coupling of this aldehyde 9 with phosphorane 6 provides 10 in 45% yield. The dimethyl acetal 10 is deprotected using acidic conditions. Treatment 11 with phosphorane 6 produces diester 12. The ester group is also DIBAL-HStarting with alcohols and subsequently with MnO₂Oxidation to form C₁₀Dialdehyde 14. The trans stereochemistry of 14 was determined by NMR (nuclear magnetic resonance) data. In particular, C of the compound₂Symmetry provides¹³5 desired resonances in the C nuclear magnetic resonance spectrum, and¹the H NMR spectrum showed signals at δ 9.54(1H), 7.07(2H) and 1.95 (3H).

g.CH₂Cl₂45 percent; h. macroporous resin 15, H₂42 to 65 percent of O and acetone; i.6, CH₂Cl₂50% -81%; DIBAL-Hhexane, 75% -81%; k.MnO₂26 to 58 percent of acetone.

The yield range in steps h to k reflects the improvement in the separation from the initial pilot study to the scale reaction.

Tiglic acid 15 is converted to phosphorane 20 in a sequence of 4 steps. Fisher esterification of 15 provides methyl ester 16. Reaction with NBS provided a mixture of 59% methyl γ -bromotiglate, and 26% methyl α -bromotiglate, with the remainder being unreacted starting material. Based on reported literature⁵It is expected that regioisomers will form. In the following step, the α/γ mixture of phosphonium salts is recrystallized to provide the desired γ -phosphonium bromide 19⁽⁶⁾. Subsequent treatment with sodium hydroxide affords the phosphorane 20.

I.H₂SO₄MeOH, 42%; nbs, benzoyl peroxide, 59%; TPP, C₆H₆，40％；o.NaOH、H₂O，81％。

Phosphoranes 20 and C₁₀Dialdehydes 14 are coupled to one another by refluxing in benzene⁶. Dimethyl crocinate 21 is isolated as a red powder. Saponification of the methyl ester proved to be much more difficult than expected. THF/H with 2eq (eq) NaOH₂The ester 21 was treated and refluxed at room temperature with O solution without changing the starting material. Solubility appears to be a significant problem, and thus pyridine is added. This did dissolve most of the solids, but refluxing a mixture of pyridine and 2.5N NaOH did not produce the product. Standard THF/2.5N NaOH saponification conditions also had no effect on the esters. Finally, refluxing overnight with 40% NaOH/methanol proved successful, yielding TSC 1 as an orange solid.

p.C₆H₆Refluxing, 33% -38%; q.MeOH, 40% NaOH aqueous solution (aq.NaOH), 58% -65%.

Attempt to dissolve the TSC to obtain¹H nuclear magnetic resonance spectroscopy. However, TSC is virtually insoluble in most common organic solvents (chloroform, DMSO, pyridine, methanol, acetone, and glacial acetic acid). TSCs prepared from this procedure were characterized by Infrared (IR), Ultraviolet (UV), High Pressure Liquid Chromatography (HPLC) and elemental analysis. IR shows at 1544cm^-1And 1402cm^-1The characteristic absorbance of (corresponding to the conjugated carboxylate). UV and HPLC are consistent with authentic TSC⁷. Elemental analysis gave satisfactory values.

The overall yield of this reaction sequence was 1.5% (based on furan).

The synthesis is detailed below:

all reagents and chemicals were purchased from Aldrich or Sigma and used as received unless otherwise indicated. Solvents were purchased from Fisher Scientific as ACS testReagent or HPLC grade and used without further purification. Anhydrous solvent with Sure/Seal^TMBottled form was purchased from Aldrich and used directly without further purification. Deionized water was obtained from an indoor Culligan water treatment system.

The melting points obtained on Mel-Temp II were not corrected. The infrared spectrum was measured on a Perkin-Elmer 1600FTIR spectrophotometer. Nuclear magnetic spectra were measured on a JEOL FX90Q spectrometer using a 5mm multinuclear probe with internal or external deuterium locks depending on the characteristics of the sample. Proton and carbon nuclear magnetic resonance chemical shifts are specified relative to TMS or deuterated solvents, respectively. Phosphorus nuclear magnetic resonance spectroscopy is typically performed in a proton decoupling mode with a coaxial insert tube of 5% aqueous phosphoric acid as an external standard.

Routine analysis was performed by gas chromatography on a Varian 3700 gas chromatograph equipped with a flame ionization detector and a Hewlett Packard 3394A integrator to assess reaction progress or estimate product composition. 1 microliter of the solution was injected with helium carrier gas onto a 15 meter DB5 column (0.53mmID and 1.5 μm film thickness), using a temperature program, increasing the temperature from 50 ℃ to 250 ℃ at 20 ℃/min and holding at 250 ℃ for 10 minutes. Typically, the temperature of the injector and detector is typically set at 250 ℃.

Thin layer chromatography was performed on a Baker-flex 2.5cm by 7.5cm silica gel plate with or without a fluorescent indicator (1B2 or 1B2-F) depending on the method of detection. The components on the spreader plate were detected by ultraviolet light.

Elemental analysis was performed by Quantitative Technologies, inc.

[ (carbethoxy) methylene group]Triphenylphosphine (4)²

(ACL-G29-1)

Triphenylphosphine (235.6g, 0.90mol) was dissolved in EtOAc (540 ml). All solids needed approximately 30 minutes to dissolve. The process is endothermic (when the ambient temperature is 20 ℃, the solution is cooled to 13 ℃). A solution of ethyl bromoacetate (100ml, 0.90mol) in EtOAc (400ml) was added dropwise over a period of 1.5 hours. A white precipitate formed during the addition. Stir at ambient temperature (18 ℃) overnight (20 h).

By using large amounts of Et₂The solid was collected by vacuum filtration rinsing. Drying overnight in vacuo at 45 ℃ gave 356.3g of 3 as a white solid in 92.6% yield (0.83 mol).¹The H nuclear magnetic resonance is consistent with the value reported in the literature.

The solid was dissolved in dichloromethane (3L) and treated with 1M NaOH (3.6L) in a 12L flask with vigorous shaking for 45 minutes. The organic layer was separated and the aqueous phase was extracted with additional dichloromethane (2X 1L). The organic layer was dried (MgSO₄) And concentrated until an amount of about 1L remains. Taking out a small amount of the substance and passing¹H nuclear magnetic resonance detection is carried out, and the result is found to be consistent with the value reported in the literature.

[1- (ethoxycarbonyl) ethylene group]Triphenylphosphonium iodide (5)²

(ACL-G29-2)

When the reaction flask was cooled in an ice bath, the material from ACL-G29-1 was treated with methyl iodide (64.0ml, 1.03 mol). When the addition was complete (1h), by TLC (silica gel, 10% MeOH/CHCl)₃) The reaction mixture was examined and showed that a considerable amount of the starting material remained. The ice bath was removed and the reaction mixture was checked by TLC after 1.5h, based on the consistency of the main band (the starting band (s.m. streamed)) and the reaction appeared to be complete. The reaction mixture was concentrated on a rotary evaporator and when most of the solvent was removed, the product started to foam and spread to the steam tube. The resulting phosphonium salt 5 is a very viscous slurry which is kept as a solution in methylene chloride to facilitate handling. Due to the nature of 5, the material was not quantified.

[1- (ethoxycarbonyl) ethylene group]Triphenylphosphine (6)²

(ACL-G29-2A)

A part of 5 is dissolved in CH₂Cl₂(350ml) and stirred vigorously with 1M NaOH (500ml) for 45 minutes. The organic layer is separated off with CH₂Cl₂The aqueous phase was extracted (2X 100 ml). The combined organic layers were dried (MgSO)₄) And concentrated to yield 8.0g of yellow solid 6.¹The H nuclear magnetic resonance spectrum is consistent with the value reported in the literature.

Fumaric aldehyde bis (dimethyl acetal) (8)³

(ACL-G29-3)

A solution of anhydrous MeOH (650ml) and furan (88.0g, 1.29mol) was cooled to-45 ℃ under nitrogen. Bromine solution (68.0ml, 1.32mol) was added dropwise over a period of 2.5 hours at a rate to maintain ≦ -45 ℃. The red colored solution was allowed to rise to-10 ℃ over a 2.5 hour period and held for an additional 2 hours. The reaction mixture was light amber in color. 5g of Na was added₂CO₃A considerable amount of evolved gas and an exotherm of 4c were generated. The reaction mixture was cooled with dry ice and the remaining Na was removed₂CO₃(total 210g) was added over a 50 minute period. After holding at-10 ℃ overnight (11 hours), the cold bath was removed and the reaction mixture was allowed to warm to room temperature and stirred for 20 hours.

The salts were removed by vacuum filtration and the filtrate was vacuum distilled using a Vigreux column until approximately 150ml of filtrate was removed. Additional salt is precipitated and causes the distillation tank to boil violently. After filtration, an additional 150ml of additional salt was distilled and more precipitated from the solution. Again, severe bumping is a problem. Cooling the still, filtering, and adding Et₂The filtrate was treated with O (400ml) and the precipitate was removed by vacuum filtration. At least 120g of salt was collected (early produced salt was discarded without quantification). Most Et was aspirated at 25 ℃ with a water aspirator₂O was removed on a rotary evaporator. The distillation was repeated using a Vigreux column and 175.2g (76.9% yield) of clear, colorless liquid 8 was collected with a boiling point of 86-92 ℃/9 torr (85-90 ℃/15 torr literature).¹The H nmr spectrum is consistent with the desired product. GC analysis: 81.9% pure.

Fumaric aldehyde mono (dimethyl acetal))(9)⁴

(ACL-G29-4)

Fumaric aldehyde bis (dimethylacetal) 8(5.29g, 0.03mol) was dissolved in acetone (120 ml). Sequential addition of H₂O (1.80ml) and macroporous resin 15(1.20 g). The mixture was stirred vigorously for 5 minutes and then filtered to remove the resin. During this time, the solution turned from colorless to yellow. The filtrate was concentrated on a rotary evaporator at room temperature and the light brown residue was distilled on a kugelrohr (37 ℃/200 mtorr) to yield 2.80g of yellow liquid 9 in 71.8% yield. When bumping occurred in the first distillation still, a small amount of material was lost.¹The H nmr spectrum was consistent with the expected product and indicated 80% pure by GC analysis.

(ACL-G29-7)

Fumaric aldehyde bis (dimethylacetal) 8(72.1g, 0.41mol) was dissolved in acetone (1600 ml). Addition of H₂O (25.0ml) and macroporous resin 15(16.7g, pre-washed with acetone). The mixture was stirred vigorously for 5 minutes and then filtered to remove the acidic resin. The reaction mixture was light yellow in color, much lighter than the previous large scale preparation. GC analysis showed 34.5% product and 46.1% starting material. The resin was treated for an additional 5 minutes. GC analysis showed 59.5% product and 21.7% starting material. The resin was treated for a further 10 minutes (total time 20 minutes). GC analysis showed 73.9% product and 2.0% starting material. The filtrate was concentrated on a rotary evaporator at room temperature to provide 54g of a brown oil. Vacuum distillation produced a yellow-green oil, 34.48 g. GC analysis showed 64.7% pure (8.22 min) with 17.5% (9.00 min) and 6.9% (9.14 min) of the major impurities. The net recovery yield was 22.3g (0.17 mol). Analysis of the front cut by GC showed a very unclean material.

(ACL-G29-13)

Macroporous resin 15(8.61g) was stirred in acetone (100ml) for 30 minutes and collected by filtration. Acetal 8(35.0g, 0.16mol) was dissolved in acetonitrile (620ml) with mechanical stirring, acidic resin added and deionisedWater (10.0ml, 0.55 mol). By TLC (10: 3, n-hexane: Et)₂O) the progress of the reaction was monitored and after 15 minutes most of the starting material had been converted. After 20 minutes, only traces of dimethylacetal were detected. The resin is removed by filtration and the filtrate is concentrated on a rotary evaporator at a temperature of 40 ℃ or less. The crude product was loaded onto a Biotage column (7.5X 9.0cm) using a column containing 15% Et₂The elution of O in hexane gave 19.8g of product in 65% yield.

6, 6-dimethoxy-2-methylhexane-2, 4-dienoic acid ester (10)²

(ACL-G29-5)

Ylide 6(7.80g, 22mmol) was dissolved in dichloromethane (65 ml). A solution of fumaric aldehyde mono (dimethyl acetal) 9(2.80g, 17mmol) was added and the solution was stirred overnight. The solvent was removed on a rotary evaporator under reduced pressure. Of the crude product¹H nmr indicated the presence of the desired product. Crystals (presumably triphenylphosphine oxide) formed upon standing. The solid (14.1 g after drying by vacuum filtration) was slurried in petroleum ether and filtered. The filtrate was concentrated to give a yellow oil with a solid precipitate, which was dissolved in dichloromethane (15ml) and chromatographed on a Biotage 4cm x 7.5cm column, eluting with dichloromethane to give 1.8g of yellow oil 10 in 50% yield. Of the yellow oil¹The H NMR spectrum corresponds to that reported in the literature, but traces of dichloromethane (0.75eq) remain, so the material is placed on a rotary evaporator for 45 minutes. The mass was reduced to 1.5g, yield 40.6%, and dichloromethane resonance disappeared. The major peak of the GC analysis occurred at 12.6 minutes, 87.5% (50 ℃, 5min hold, 20 ℃/min ramp to a final temperature of 250 ℃).

(ACL-G29-6)

A solution of ylide 6(59.2g, 0.16mmol) was dissolved in dichloromethane (650ml), the solution was cooled in an ice bath and a solution of 9(25.7g, 0.19mol) was added. The solution was stirred overnight to allow the ice bath to melt. TLC (n-hexane: Et)₂O, 10: 3) showed that at least 3 other compounds were very close to the product after spreading. The result of detecting the aldehyde by GC analysis showed 50% purity. The solvent was removed to yield a solid/oil mixture.

(ACL-G29-8)

Ylide 6(59.2g, 0.16mmol) and acetal 9(0.19mol) were coupled in dichloromethane (1.1L) and treated as above to give 80g of a yellow-green oil. A portion of the crude reaction mixture (4.13 g of the initial 80 g) was placed on a Kokugelrohr and distilled at 50 deg.C/250 mTorr. Concentration gave 2.28g of a colourless oil,¹h NMR indicated it to be the initial aldehyde, while 1.85g of product 10 remained in the still. A Kugelrohr distillation was carried out at 50 ℃ at 200 mTorr to remove volatile constituents (35 g dry weight) from the bulk of the crude product.

2-methyl-6-oxo-hexane-2, 4-dienoic acid ethyl ester (11)²

(ACL-G29-9)

Acetal 10(ACL-G29-8, 1.85G, 9mmol) from the experimental still was dissolved in acetone (33 ml). Deionized water (0.50ml) and macroporous resin 15(0.35g, pre-washed with acetone) were added. The mixture was stirred for 20 minutes. Filtered and concentrated on a rotary evaporator to yield 1.53g of a yellow-green oil. Chromatography was performed on a Biotage column of 4.5 cm. times.7 cm and containing 15% Et₂Hexane elution of O was performed. This system provided incomplete separation, but 0.32g of the major product was isolated and analyzed;¹h NMR spectra were consistent with literature values, IR (1711, 1682 cm)^-1) Consistent with the desired product. And GC 95.6%. An additional 0.35g of product is recovered, but more or less doped with polar substances.¹H-nmr spectra showed a fairly clean material. GC 90.6%. Yield: 42 percent.

2, 7-Dimethylocta-2, 4, 6-triene-1, 8-dioic acid diethyl ester (12)²

(ACL-G29-10)

Aldehyde 11(0.65G, 3.5mmol) from G29-9 was dissolved in dichloromethane and magnetically stirred. The ylide (1.59g, 4.4mmol) was added. The yellowish green solution turned dark yellow within a few minutes. TLC was performed after 10 min, which indicated almost complete consumption of the starting material. After stirring for 20 hours, the reaction mixture (brown solution) was filtered through a pipette partially filled with silica gel. The filtrate was concentrated to give a brown solid. With small amounts of CHCl₃The solid was dissolved in 5% Et₂O in hexane. Chromatography was performed on a Biotage column of 4 cm. times.7.5 cm, using 5% Et₂Hexane elution of O was performed. 0.45g of the main product was isolated as a white crystalline solid in 50% yield.¹The H nuclear magnetic resonance spectrum is consistent with data reported in the literature.

(ACL-G29-14)

After chromatographic purification, an additional amount of 12, 21.8g, was prepared as described above in 81.6% yield.¹The H nuclear magnetic resonance spectrum is consistent with the required product.

2, 7-Dimethylocta-2, 4, 6-triene-1, 8-diol (13)²

(ACL-G29-11)

Diester 12(0.45g, 1.8mmol) was dissolved in anhydrous hexane (15.0 ml). It appeared as if some of the material had dissolved, but the mixture was very cloudy. When the mixture was cooled in a-78 ℃ bath, more material began to show up from the solution. Pure DIBAL-H (2.50ml) was dissolved in anhydrous hexane (total volume 10.0ml) and a portion (approximately 2ml) of the diester was unnoticeably siphoned into the reaction mixture when the diester was cooled in a dry ice bath. An additional amount of DIBAL-H solution was added until the total amount added reached 5.0ml (6.7 mmol). Introducing the CO into a reactor₂The bath is warmed up. After stirring for 2 hours 50 minutes, TLC indicated complete consumption of the diester. The bath temperature was adjusted to-20 ℃ and allowed to warm to 0 ℃ in 20 minutes. By H₂The O/silica gel (2ml/7g) mixture was treated for 30 minutes. Adding K₂CO₃And MgSO₄. Filtering to removeThe solid was rinsed thoroughly with dichloromethane. Concentration was performed to yield 0.14g of a white solid in 50% yield. Note: TLC R_f＝0.21(5％MeOH/CHCl₃) The polarity is strong. Rinsing with dichloromethane may not be sufficient to recover the entire product.¹The H nuclear magnetic resonance spectrum is consistent with the value reported in the literature.

(ACL-G29-15)

The diester (5.4g, 21mmol) was dissolved in anhydrous hexane (175ml, low solubility), cooled in a-78 ℃ bath and treated with DIBAL-H solution (14.5ml, 50ml in anhydrous hexane) for 35 min. Vigorous gas generation was observed during the addition. The color of the resulting slurry initially changed from white to dark yellow, and the sample became lighter when additional DIBAL-H was added. The temperature was allowed to rise to-40 ℃ over 2 hours and then transferred to a-28 ℃ bath overnight. By H₂A homogeneous mixture of O/silica gel (4ml/14.4g) was treated for 30 minutes. Adding MgSO₄(7.5g) and K₂CO₃(5.1g), and the reaction mixture was removed from the cooling bath. Stirred for 20 minutes and then filtered in a sintered glass funnel. The resulting solid was washed with dichloromethane, which resulted in the formation of a considerable amount of precipitate. The temperature was increased while placing on the rotary evaporator to dissolve the precipitated solid. The solid remaining on the sintered glass funnel was washed with EtOAc (4X 75mL) and the filtrate was concentrated.

CH₂Cl₂Rinsing provided 1.7g of a pale yellow solid,¹h nuclear magnetic resonance is consistent with the reported value in the literature; EtOAc bleaching provided 1.0g of a milky white solid,¹h nuclear magnetic resonance is consistent with the reported value in the literature; a total of 2.7g was recovered in 75% yield.

(ACL-G29-17)

In N₂The diester (16.4g, 6.5mmol) was stirred in anhydrous hexane (500ml) and cooled to-78 ℃. A solution of DIBAL-H (45ml, 253mmol) in hexane (150ml) was added over a period of 1 hour. The temperature was allowed to rise to-30 ℃ and stirred overnight (total time 17.5 hours). Addition of H₂Homogeneous mixture of O/silica gel (12.3g/43.7g), handThe mixture was vortexed for 45 minutes. Adding K₂CO₃(15.5g) and MgSO₄(23.5 g). Vortex for another 30 minutes. Filtration was performed in a sintered glass funnel, rinsing with dichloromethane (to form ppt grade, presumably due to evaporative cooling) was performed, and the filtrate was concentrated. The solid was washed several times with EtOAc (ca 100ml one part, 2L total volume) and combined with the original filtrate. Concentration gave 8.9g of a yellow solid in 81% crude yield.¹The H nuclear magnetic resonance spectrum is consistent with the required product.

2, 7-Dimethyloctyl-2, 4, 6-triene-1, 8-dialdehyde (14)²

(ACL-G29-12)

In N₂MnO Cooling in an Ice bath₂(7.80g, 90mmol) of slurry. A solution of diol 13(0.14g, 0.8mmol) was added as an acetone solution (5.0ml) by pipette. An additional 2.0ml of acetone was used to rinse the flask and complete the transfer. The reaction mixture was stirred while allowing the ice bath to melt overnight. The solid was removed by filtration over Hyflo and concentrated to give a yellow solid. With a minimum of CHCl₃This material was dissolved in 10% Et₂O/hexane, and applied to a silica gel column (30 mm. times.190 mm) and washed with 10% Et₂O/hexane elution. When the product was eluted, it could be followed as a yellow band, isolating 37mg of 14 as a pale yellow solid in 26% yield.¹The H nuclear magnetic resonance spectrum is consistent with the value reported in the literature.

(ACL-G29-16)

In N₂A solution of diol 13(2.70g, 16mmol) in acetone (500ml) was cooled in an ice bath and MnO was added in portions over a 20 minute period₂(60.0g, 0.69 mol. when the reaction mixture was stirred overnight, the ice bath was allowed to melt the reaction mixture was filtered through Hyflo and the filtrate was concentrated to give 1.6g of a yellow solid in crude yield of 61%.¹The H nuclear magnetic resonance spectrum is consistent with the value reported in the literature. The crude yellow solid was dissolved in dichloromethane (with the addition of a small amount of 10% Et₂Hexane of O) and loaded into 4 × 7.5cm Biotage siliconAnd (5) gluing the column. Eluting with 10% Ether in hexane (1L) and then with a polar-enhancing solution containing 15% Et₂Ohexane (1L) and 20% Et in₂Hexane (0.5L) of O. 1.0g of a yellow solid was recovered in 38% yield.¹The H nuclear magnetic resonance spectrum is consistent with the required product.

(ACL-G29-21)

In N₂A solution of diol (9.31g, 60mmol) dissolved in acetone (500ml) was cooled in an ice bath. Adding MnO₂(100g, 1.15mol) and the reaction mixture was stirred overnight while allowing the ice bath to melt. After 24 hours, by infrared detection, it was found that a significant amount of product was formed, but still considerable alcohol was present. An additional 50g of oxidant was added and stirring continued overnight. Filtering a portion of the reaction mixture and passing¹H nmr was detected and the reaction appeared to have completed based on the consumption of starting material. The remaining reaction mixture was filtered through a Hyflo pad and rinsed thoroughly with acetone. Concentrate to give a dark yellow solid. Azeotroped with 40ml of benzene and dried in vacuo at 40 ℃ for 5 hours and then at room temperature overnight. The recovery was 5.28g, 58% yield.¹The H NMR and IR spectra were consistent with the desired product.

Tiglic acid methyl ester (16)

In a 2L three-necked flask equipped with an overhead stirrer, condenser and thermometer, a solution of tiglic acid 15(89.8 g; 0.9mol) and 5ml of concentrated sulfuric acid (0.09mol) dissolved in 900ml of methanol was heated under reflux for 20 hours. The solution was cooled to 25 ℃ and stripped of excess methanol on a rotary evaporator at 30 ℃ and 27in Hg vacuum. GLC analysis of the recovered methanol distillate indicated that the product was located in the overhead (overheads). The resulting two-phase, light-brown concentrate is dissolved in 500ml of diethyl ether and washed successively with 250ml of water, 250ml of 10% aqueous sodium bicarbonate solution and 250ml of saturated brine. The ether solution was dried over anhydrous potassium carbonate, filtered and stripped on a rotary evaporator at 25 ℃ under 17in Hg vacuum to yield 43.6g (42% yield) of crude methyl tiglate as a nearly colorless oil. GLC analysis showsOne of the major volatile products, retention time was 2.7 minutes relative to the initial 3.8 minutes retention time of tiglic acid. In CDCl₃The proton nmr carried out in (c) showed the expected signal with trace ether contamination: 1.79ppm (d, 3H), 1.83(s, 3H), 3.73(s, 3H), 6.86(q, 6.6 Hz). IR (pure KBr): at 1718cm^-1Ester carbonyl group of (a). The use of this oil is the same as in the next step.

Gamma-Brovigonic acid methyl ester (17)⁵

A stirred mixture of crude methyl tiglate (43.6 g; 0.38mol), N-bromosuccinimide (68 g; 0.38mol) and 70% benzoyl peroxide (5.34 g; 0.015mol) dissolved in 500ml of carbon tetrachloride was heated under reflux for two hours in a 1L four-necked flask equipped with an overhead stirrer, thermometer and condenser. After cooling to 20 ℃ the insoluble succinimide (38.1g, 100% recovery) was suction filtered off. The filtrate was washed 3 times with 250ml water, over MgSO₄Drying and then gas-extracting at 25 ℃ under 26in Hg vacuum on a rotary evaporator gave 78.8g of a yellow oil. In CDCl₃The proton nmr performed on this oil produces a complex nmr spectrum. The methylene proton for the desired γ -bromo ester was assigned a bimodal peak centered at 4.04ppm (8.6Hz), while the same proton for the α -bromo isomer was assigned a single peak at 4.24 ppm. The proton integration of these signals and methyl multiplets from 1.6ppm to 2.0ppm imply the following composition (mol%):

gamma-bromo ester: 59 percent of

α -bromo ester: 26 percent of

Starting materials: 15 percent of

This crude oil was used in the next step without any purification.

This reaction was also carried out on a 0.05mol scale using only 0.87 equivalents of N-bromosuccinimide, under otherwise identical conditions. Based on its proton nuclear magnetic resonance spectrum, the composition of this crude oil was estimated to be 52% gamma-bromo ester, 24% alpha-bromo ester and 23% unreacted methyl tiglate. GLC analysis of this oil was slightly more complex, showing other minor components.

Triphenylphosphine onium salt of gamma-methyl tiglate (19)⁶

A stirred solution of crude methyl γ -bromotiglate (78.8g) dissolved in 350ml of benzene was treated by dropwise addition of triphenylphosphine (95 g; 0.36mol) dissolved in 350ml of benzene in a 2L four-necked flask equipped with a thermometer, a 100ml constant pressure addition funnel and a condenser connected to a static nitrogen system over a period of 1.75 hours. Under other ambient conditions, the temperature of the mixture rose slightly exothermically from 24 ℃ to 27 ℃. After addition, the reaction mixture was stirred vigorously overnight to provide a slurry of white solid containing a yellowish gum adhered to the flask walls. The white solid was suction filtered onto a sintered glass funnel without breaking the yellowish gum. The flask was washed twice with 100ml benzene and the washings were poured onto the filter. The filter cake was washed with 50ml of benzene and then twice with 50ml of hexane. The wet cake was dried in a vacuum oven at ambient temperature for 5.5 hours. The dried white powder [93 g; mp 125 ℃ (decomposed) was dissolved in 150ml acetonitrile to give a clear yellow solution. 300ml of ethyl acetate were added to this hot solution and the product started to crystallize after about 100ml of ethyl acetate had been added. The flask was kept in a refrigerator overnight. The product is filtered off with suction and washed with a minimum of acetonitrile and ethyl acetate solution in a ratio of 1: 2; 45.0g, mp 187-190 ℃ (decomposed), and mp 183 ℃ (decomposed).

The sticky solid in the reaction flask was recrystallized from 10ml acetonitrile and 20ml ethyl acetate. Also, additional solids precipitated from the benzene mother liquor overnight. These solids were filtered and recrystallized in the same manner. Both samples were frozen for 2 hours and suction filtered to give an additional 13.3g of product.

The benzene filtrate was stripped on a rotary evaporator, the yellow oil was dissolved in 10ml of acetonitrile and precipitated with 20ml of ethyl acetate.The slurry was kept in a refrigerator overnight to give additional product as a white solid; 4.6g, m.p.185-187 deg.C (decomposition). The total yield of the phosphonium salt required as a white solid was 62.9g or 36.2% yield based on crude methyl tiglate. Proton nuclear magnetic resonance (CDCl)₃TMS) ppm 1.55(d, 4Hz, 3H), 3.57(s, 3H), 4.9(dd, 15.8 and 7.9Hz, 2H), 6.55 (width q, 6.6-7.9Hz, 1H), 7.4-7.9(m, 15H). Proton decoupled phosphorus nuclear magnetic resonance (CDCl)₃，5％H₃PO₄Aqueous coaxial external standard) 22.08 ppm. Local carbon nuclear magnetic resonance (CDCl)₃)：CO₂CH₃(166.6ppm，d，J_CP3Hz), olefinic CH (117.5ppm, d, J_CP＝86.1Hz)，CO₂CH₃(52.0ppm)，Ph₃P-CH₂(25.4ppm，d，J_CP50.6Hz) and CH₃(13.4ppm，d，J_CP2.4 Hz). Partial IR (KBr sheet): ester carbonyl at 1711cm^-1To (3).

[ 3-carbomethoxy-2-buten-1-ylidene group]Triphenylphosphine (20)⁶

In a 5L five-necked flask equipped with an overhead stirrer, addition funnel and thermometer, a solution of sodium hydroxide (5.12 g; 0.128mol) dissolved in 250ml of water is added dropwise over a period of 41 minutes at 25 ℃ to a vigorously stirred solution of the triphenylphosphine salt of methyl γ -tiglate (58.3 g; 0.128mol) dissolved in 2,500ml of water. The yellow slurry was stirred at room temperature for 10 minutes and then suction filtered. The filter cake was washed with 1,800ml of water and then dried thoroughly on the filter with a nitrogen blanket. The yellow solid was then dried in a vacuum desiccator at room temperature under 27 "Hg vacuum at P₂O₅The mixture was dried overnight to obtain 35.3g (73.7% yield). mp-145 ℃ -150 ℃. The literature mp ═ 145 ℃ to 165 ℃. In CDCl₃The proton decoupled phosphorus NMR performed in (1) showed two peaks at 17.1ppm and 21.1ppm, with a ratio of 93: 7. Proton nuclear magnetic resonance (CDCl)₃TMS) ppm 1.89(s, 3H), 3.58(s, 3H), 7.3-7.8(m, 17H). A small but detectable single peak, attributable to impurities, was also seen at 1.74ppm in this NMR spectrum.This solid was used in the next step without purification.

Crocetin dimethyl ester (21)⁶

(ACL-G29-18)

Dialdehyde 14(0.48g, 2.9mmol) was added to a 100ml round bottom flask. 20ml of benzene was added and the solid was dissolved by magnetic stirring. The ylide was added and the compound was washed into the flask with an additional 10ml of benzene. The temperature was increased and vigorous reflux was carried out for 6 hours. The reaction mixture was cooled overnight. In contrast to literature reports, very small amounts of solids are formed. The reaction mixture was concentrated and the residue was dissolved in 30ml MeOH and boiled for 30 min. Upon cooling to room temperature, the solid was collected by vacuum filtration. By dissolving 20mg of the solid in 0.5ml CDCl₃The nmr samples were prepared, and it was somewhat surprising that this necessitated heating with a heat gun to completely dissolve. Recording¹H nuclear magnetic resonance spectrum and the resonance spectrum was found to be consistent with the desired product. The remaining material was dissolved in hot benzene and filtered, the filtrate was concentrated and dissolved in MeOH, cooled in an ice bath and 334mg of red solid was collected in 33% yield. This material did not appear to be more soluble than the initially isolated material.

(ACL-G29-18A)

In N₂Dialdehyde 14(5.78g, 35mmol) was dissolved in 300ml of benzene. The ylide 20(35.3g, 94mmol) was added and the mixture was refluxed for 6 hours at elevated temperature to form a dark red solution. After the reaction mixture was cooled overnight, the red solid was collected by vacuum filtration and rinsed with methanol. It was transferred to 500ml RBF and refluxed with about 65ml methanol for 30 min. Cool and collect the red solid. Rinsed with cold methanol and dried in vacuo to give 3.00g of a red solid, 21.¹H NMR and IR spectra were consistent with the desired product.

The initial filtrate (from the reaction mixture) was concentrated on a rotary evaporator and the dark residue was dissolved in 100ml of methanol and refluxed for 40 minutes. Cool in an ice bath and collect the red solid by vacuum filtration.Rinsed with cold methanol and dried in vacuo to yield 1.31g of a red solid, 21.¹The H nuclear magnetic resonance spectrum is consistent with the required product.

The filtrates were combined, concentrated, and dissolved in 75ml of methanol and left overnight at room temperature. The red solid was recovered by vacuum filtration: 0.38 g.¹The H nuclear magnetic resonance spectrum is consistent with the required product.

More solids were formed in the filtrate. Separation was performed by vacuum filtration to give 0.127g of a red solid. The IR results are consistent from the foregoing. Total recovery: 4.89g, 39% yield.

Saponification attempts with THF/NaOH

(ACL-G29-19)

A stirred suspension of diester 21(100mg, 0.28mmol) in THF (2ml) and 1N NaOH (0.56ml, 2eq) was added. Stir at room temperature overnight. TLC showed only the starting material. The reflux was carried out at elevated temperature and after several hours no change occurred. 6ml of THF was added in an attempt to dissolve more of the solid, but this did not appear to work. Reflux was continued overnight. More THF (about 6ml, TLC showed only starting material) was added and refluxed overnight. Concentrating and passing through¹H nmr was detected and the results showed only the starting material (based on a combination of methyl and methyl esters). Dissolved in 10ml of pyridine, and the temperature was raised in a heating mantle. 2.5N NaOH (1.0ml) was added. The dark orange solution turned dark red after a few minutes. The heating mantle was removed and a solid began to form, and the mantle was reapplied for 30 minutes and then stirred at room temperature overnight. Concentration was carried out in a high vacuum. The residue was insoluble in chloroform, DMSO, pyridine, slightly soluble in water. IR (paraffin paste) showed C ═ O absorption characteristics of the starting material.

Saponification with 2.5N NaOH and THF

(ACL-G29-20)

Diester 21(37mg, 0.10mmol) was weighed into a flask and stirred in 4ml of diethyl ether. Solids still remained when the solvent was orange. 1ml of 2.5N NaOH was added and the temperature was raised to reflux. After half an hour, most of the ether had evaporated. The ether was replaced with THF (3ml) and refluxing was continued for several hours. The solid was collected by vacuum filtration, rinsed with deionized water and then dried in a vacuum oven. IR only shows the starting material.

Saponification with 40% NaOH (1)

(ACL-G29-22)

Diester 21(32mg, 8.9mmol) was weighed into a flask and stirred in methanol (1.5 ml). Solids still exist when the solvent is orange/red. 1.5ml of 40% NaOH was added and the temperature was raised to reflux for 17 hours. After cooling to room temperature, an orange solid was collected by vacuum filtration and rinsed with deionized water. Drying was carried out in vacuo at 40 ℃ to give 21mg of 1 as an orange powder in 59% yield. IR (KBr piece) 3412cm^-1，1544cm^-1，1402cm^-1The compound may be hygroscopic, with high magnetic field carbonyl shifts consistent with conjugation.

(ACL-G29-22A)

Reflux was repeated with 35mg of diester 1 for 15 hours. The reaction mixture was cooled in an ice bath, collected by vacuum filtration and washed with chilled deionized water. Drying was carried out in vacuo at 40 ℃. 25.5mg of 1 as an orange solid were recovered in 65% yield.

(ACL-G29-23)

Diester 21(0.48g, 1.3mmol) was dissolved in methanol (15.0ml) and 40% sodium hydroxide (15.0ml) and heated to reflux. After about 2 hours, the heterogeneous red mixture turned orange. After 6 hours the heating was stopped and the mixture was allowed to cool overnight. The orange solid was collected by vacuum filtration and washed with chilled deionized water. Drying in vacuo gave 0.36g of a fluffy orange solid in 68% yield.

(ACL-G29-24)

Diester 21(1.10g, 3.1mmol) was placed in a 100ml recovery flask and washed with methanol (20ml) and 40% NaOH (20ml)) Heating and refluxing for 12 hours. After cooling in an ice bath, an orange solid was collected by vacuum filtration and rinsed with deionized water. Drying in vacuo afforded 1.4g of product in 100% yield. C₂₀H₂₂O₄Na₂-0.4H₂Analytical calculation of O: c, 63.29; h, 6.05; na, 12.11; h₂O, 1.90. Measured value: c, 63.41; h, 6.26; na, 11.75; h₂O，1.93。

(ACL-G29-25)

Diester 21(3.00g, 8.4mmol) was refluxed in methanol (80ml) and 40% NaOH (60ml) for 12 hours. 2.7g of product was isolated as an orange solid in 80% yield as above. C₂₀H₂₂O₄Na₂-0.4H₂Analytical calculation of O: c, 63.29; h, 6.05; na, 12.11; h₂O, 1.90. Measured value: c, 63.20; h, 6.00; na, 11.93; h₂O, 1.81. Samples ACL-G29-23, ACL-G29-24, and ACL-G29-25 were ground on an agate mortar and combined into ACL-G29-A.

Reference to the literature

E.buchta and f.andree naturwiss.1959, 46, 74;

2.F.J.H.M.Jansen，M.Kwestro，D.Schmitt，J.Lugtenburg Recl.Trav.Chim.Pays-Bas 1994，113，552；

3.R.Gree，H.Tourbah，R.Carrie Tetrahedron Letters 1986，27，4983；

4.G.M.Coppola Syn.Commun.1984，1021；

d.s.letham and h.young Phytochemistry 1971, 10, 2077;

e.buchta and f.andree chem.be.1960, 93, 1349.

Example 2

Synthesis of potassium trans norbixin

By combining C of symmetrical type containing conjugated carbon-carbon double bonds₂₀Dialdehydes and [1- (carbethoxy) methylene]And (3) performing phase coupling on the triphenyl phosphine to synthesize the trans-norbixin potassium. The preparation of this compound is similar to the preparation of sodium para-trans crocetinate previously listed, except that the starting material furan is replaced with a suitable cyclic structure. This product was then saponified with KOH/methanol solution.

Example 3

Longer synthesis of BTCS

By combining C of symmetrical type containing conjugated carbon-carbon double bonds₁₀Dialdehyde is added to excess [ 3-carbomethoxy-2-buten-1-ylidene ]]Triphenylphosphine. The preparation of this compound is similar to the preparation of sodium para-trans crocetinate previously listed, except that the starting material furan is replaced with a suitable cyclic structure. The trans 40-carbon product is then isolated using procedures such as chromatography. This product was then saponified with NaOH/methanol solution.

Example 4

Administration of TSC by inhalation

TSC was administered to rats via the inhalation route. TSC was directly delivered to the lungs of 10 rats. This was done by inserting a tube into the trachea and nebulizing 0.2ml of TSC solution (TSC dissolved in dilute sodium carbonate solution) with about 3-6 ml of air. For all study doses (0.5 mg/kg-2 mg/kg), about 20% of the drug was present in the bloodstream within 1 minute after administration. For doses of 0.8mg/kg to 1.6mg/kg, the drug is present in the bloodstream for at least 2 hours.

Example 5

Improved synthesis method

Preparation of 2-butenyl-1, 4-diphosphonic acid tetraethyl ester

A250 ml three-necked flask was equipped with a Teflon coated thermocouple, a 60ml constant pressure addition funnel, and a simple distillation head. Pure triethyl phosphite (59 ml; 0.344mol) was heated at 140 ℃ under nitrogen with a heating mantle controlled by a JKem controller. A solution of trans-1, 4-dichloro-2-butene (26.9 g; 0.215mol) and triethyl phosphite (35 ml; 0.204mol) was added dropwise at 134 ℃ to 144 ℃ over a period of 93 minutes. The clear solution was then stored under nitrogen at 140 ℃. After 37 minutes, an aliquot (1 drop) in 1ml of ethyl acetate was analyzed by gas chromatography and showed the desired product, intermediate and two starting materials.

After 15.5 hours at 140 ℃, gas chromatography analysis of an aliquot (1 drop in 0.5ml EtOAc) showed the desired product with no detectable starting dichloride or intermediate. After 16 hours, the light yellow solution was cooled to room temperature under nitrogen. The pale yellow oil was distilled in a council distiller with a double ball receiver and the ball receiver was further cooled in a dry ice-acetone bath at 25 ℃ to 100 ℃ and 0.1 torr to 0.2 torr to obtain a colorless oil (14.8g) as a front cut. Gas chromatography showed only product in the retort. This light amber colored oil was distilled in a retort at 140 ℃ and 0.1 to 0.15 torr to give 66.45g of distillate as colorless oil (94.1% yield). Gas chromatography showed only one volatile component. GC-MS analysis showed this component to be the desired product, giving a small molecular ion peak at 328m/z and 191m/zBasic ion Peak (PO)₃Et₂Loss of) of the memory. Proton nmr is consistent with the desired product. Carbon nuclear magnetic resonance was consistent with the desired bis (phosphonate diester), showing only remote coupling to the allylic carbon (W-coupling) and normal carbon-phosphorus coupling.

The residue in the tank was 0.8g of a pale yellow oil.

Preparation of 1, 1,8, 8-tetramethoxy-2, 7-dimethyl-2, 4, 6-octatriene

A magnetically stirred mixture of trans-2-butenyl-1, 4-diphosphonic acid tetraethyl ester (3.3 g; 10.0mmol), methylglyoxal dimethyl acetal (2.6 ml; 21.5mmol) in 10ml toluene and 10ml cyclohexane was treated successively with anhydrous potassium carbonate (10.2 g; 73.8mmol) and powdered sodium hydroxide (1.25 g; 31.2mmol) under a nitrogen atmosphere. The solution immediately turned yellow. The resulting slurry was stirred at ambient temperature under nitrogen. The reaction was slowly exothermic and reached a maximum of 38 ℃ after about 25 minutes. Also, a sticky precipitate was formed, which hindered magnetic stirring. After 2.5 hours, gas chromatography of an aliquot of this yellow-orange solution (1 drop in 0.5ml of toluene) showed two starting materials and 3 other new ingredients.

Gas chromatography of an aliquot of this orange solution (1 drop in 0.5ml toluene) after 16.75 hours at ambient temperature showed only a small amount of the starting bis (phosphonic diester). The resulting orange mixture with stickies (unable to stir) was cooled in an ice bath and quenched with 100ml of 10% aqueous NaCl. The solid was dissolved in the aqueous solution by using a spatula. Extraction was then carried out with 200ml of ether: hexane in a ratio of 1: 1. The organic layer was washed with 10% aqueous NaCl solution (200ml), followed by washing with saturated brine (100 ml). Through Na₂SO₄The colorless organic layer was dried. Gas chromatography showed 3 major components with no detectable onsetThe substance bis (phosphonic diester). Thin layer chromatography showed two major spots and one minor spot. Suction filtration to remove Na₂SO₄And washed with ether. The filtrate was concentrated on a rotary evaporator at 35 ℃ to give 1.8g of a colorless oil. GC-MS analysis showed that the 3 major volatile components were isomeric products, giving a molecular ion peak at 256m/z and a basal ion peak [ (MeO) at 75m/z₂CH⁺]. The proton nmr results are consistent with mixtures of isomeric products accompanied by other unidentified impurities. The yield of the crude product was 70.3%.

Preparation of 1, 1,8, 8-tetramethoxy-2, 7-dimethyl-2, 4, 6-octatriene

A mechanically stirred mixture of trans-2-butenyl-1, 4-diphosphonic acid tetraethyl ester (63.2 g; 0.19mol), methylglyoxal dimethyl acetal (50 ml; 0.41mol) in 200ml toluene and 200ml cyclohexane was treated successively with anhydrous potassium carbonate (196 g; 1.42mol) and powdered sodium hydroxide (24.0 g; 0.60 mol). The solution immediately turned yellow. The resulting slurry was stirred at ambient temperature under nitrogen. The reaction exothermed to 61 ℃ after about 11 minutes and the stirred mixture was cooled in an ice bath to reduce the temperature to 35 ℃. After 4.7 hours at 29 ℃ to 35 ℃, gas chromatographic analysis of an aliquot (3 drops in 0.5ml toluene) showed no starting di (phosphonate). After about 5 hours, the mixture was cooled to 13 ℃ in an ice bath and 10% aqueous sodium chloride (400ml) was added as the temperature rose to 30 ℃. More 10% aqueous sodium chloride (1,500ml) was added and the mixture was extracted with 3,000ml of 1: 1 ether: hexane. The colored yellow organic layer was washed with 10% aqueous sodium chloride solution (2X 1,000ml), followed by saturated brine (1,000 ml). Through Na₂SO₄The colored yellow organic layer was dried, filtered and concentrated on a rotary evaporator at 30 ℃ to give 43.4g of a pale yellow oil. The gas chromatography showed 3 kindsA major component comprising 89% of the mixture without detectable starting di (phosphonate). TLC analysis showed one major component and 3 minor components.

Proton nmr showed the isomerate and toluene. The oil was further evaporated on a Kokuk distiller at 50 deg.C and 0.2 torr for 30 minutes to yield 31.9g of product. Proton nmr showed the isomeric di (acetal) product with no detectable toluene.

The yield was 65.5%.

Preparation of 2, 7-dimethyl-2, 4, 6-octatrienedial at higher payload

A magnetically stirred solution (9:03am) of the crude 1, 1,8, 8-tetramethoxy 2, 7-dimethyl-2, 4, 6-octatriene isomer (31.9 g; 124.4mmol) in tetrahydrofuran (160ml), water (80ml) and glacial acetic acid (320ml) was heated at 45 ℃ in a nitrogen atmosphere with a heating mantle controlled by a Jkem controller via a Teflon coated thermocouple. After about 30 minutes, the mixture exothermed to a maximum temperature of 54 ℃ and then returned to the set point of 45 ℃. After 3 hours, gas chromatography of an aliquot (3 drops in 0.5ml THF) showed some residual starting material, two major products and one minor product. The yellow reaction solution was cooled to 21 ℃ in an ice bath and then diluted with 4: 1 ether: dichloromethane (2,000 ml). Then, the mixture was successively treated with 20% aqueous NaCl solution (2,000 ml. times.2), 4: 1 aqueous 20% NaCl solution: 1M aqueous NaOH solution (2,000 ml. times.3)¹And 20% aqueous NaCl solution (1,000ml × 2) wash this solution (note:¹the first two washes apparently removed acetic acid because the pH was neutral. The third wash turned red and still basic indicating removal of by-products). By MgSO₄The yellow organic layer was dried, filtered and concentrated on a rotary evaporator to give 18.9g of a yellow solidAnd (3) a body. Gas chromatography showed one major and one minor component of the starting di (acetal). TLC analysis showed one major spot and several minor, more polar impurities. The solid was dissolved in 250ml of refluxing methanol, cooled to room temperature and then cooled in an ice bath for 1 hour. The slurry was suction filtered to give 14.15g of yellow soft needles. Gas chromatography showed a 95: 5 mixture of isomeric dialdehydes. The solid was recrystallized again from 200ml of refluxing methanol, cooled to room temperature and then cooled in a refrigerator overnight.

The slurry was suction filtered and washed with refrigerator-frozen methanol to give 11.2g of yellow needles. Gas chromatography showed a 97: 3 mixture of isomeric dialdehydes. TLC analysis showed one spot. The needle was dried in a vacuum oven at 45 ℃ for 160 minutes until the weight was constant at 10.75 g.

Uncorrected mp is 154-156 ℃. Literature reference²-mp 161-162 ℃ (note:²dictionary of Organic Compounds, 10 th edition: 2, Sept, 2002). Proton nuclear magnetic resonance and carbon nuclear magnetic resonance are consistent with the desired symmetric dialdehyde.

The two methanol filtrates from the recrystallization were combined together. The thin layer chromatography showed product and other impurities. The filtrate was concentrated and several harvests were collected as follows.

Harvesting of crops	Appearance of the product	Amt(g)		Isomer ratio
					2	Yellow powder	1.4		80∶20
3	Yellow needle-like substance	2.6		75∶25
					4	Yellow solid	4.45		46∶30

Harvests 2 and 3: the combined harvest was dissolved in 20ml of refluxing ethyl acetate, cooled to room temperature and then cooled in a freezer for 1 hour. The slurry was suction filtered and washed with refrigerator frozen ethyl acetate to give 1.95g yellow needles. Gas chromatography showed an 86: 14 isomer mixture. The solid was recrystallized from ethyl acetate (10ml) to give 1.55g of yellow needles. Gas chromatography showed a 92: 8 isomer ratio. The 3 rd recrystallization from ethyl acetate (10ml) afforded 1.25g of yellow needles. mp is 152-154 ℃. Gas chromatography showed a 96: 4 isomer ratio. Proton nmr confirmed the desired dialdehyde. GC-MS analysis was consistent with the desired dialdehyde, showing thatApparent M at 164M/z⁺Ion peak and base ion peak at 91 m/z.

The ethyl acetate filtrate was combined with the yellow solid from the methanol filtrate (harvest 4) and concentrated on a rotary evaporator to give 6.0g of a yellow solid. Gas chromatography showed a mixture of the two isomers in a ratio of 53: 34 and other impurities.

The solid was dissolved in 100ml dichloromethane and Davisil grade 643 silica gel (35.5g) was added. The mixture was stripped on a rotary evaporator at 35 ℃. The silica gel and its adsorbed material was then added to the sample injection assembly of the Biotage system, which already contained a plug of glass wool and a layer of sand. The silica gel was then covered with filter paper. The Biotage 75S column was pre-wetted with a solvent mixture having a radial pressure of 35psi and a solvent pressure of 20 psi. The column was eluted with 85: 15 n-hexane: ethyl acetate (6,000 ml). A void volume of 1,000ml was obtained including the pre-wet stage. Fractions of 250ml were collected and pooled based on thin layer chromatography analysis. These fractions were concentrated on a rotary evaporator at 35 ℃ as follows.

Fraction (b) of	Content(s) therein	Appearance of the product	Amt(g)	Note
					1	Blank space
2～3	A
					4	Trace amount (tr) A
5～10	B	Yellow solid	3.9	Product fraction
					11～18	Trace B or trace C			There is no evidence of close elution of impurities
19～20	Traces of B or C and D

Fraction 5-10: the yellow solid was slurried in n-hexane and suction filtered to give 2.5g of a pale yellow solid. Gas chromatography showed a mixture of dialdehyde isomers in a ratio of 67: 33.

96-97% of E, E, E-dialdehyde, the total yield of which is 10.75+ 1.25-12.0 g (yield 58.8%).

Isomerization of 2, 7-dimethyl-2, 4, 6-octatrienedial with p-toluenesulfinic acid

A2: 1 isomeric mixture of 2, 7-dimethyl-2, 4, 6-octatrienedial and its undesired isomer (2.5 g; 15.2mmol), 4-toluenesulfinic acid (0.35 g; 2.2mmol) and 50ml of anhydrous 1, 4-dioxane were heated to reflux under nitrogen for 15 minutes. An aliquot (7 drops) was diluted in 0.5ml of ether: dichloromethane at a ratio of 4: 1 and passed through K₂CO₃Drying is carried out. Gas chromatography showed a 91: 9 mixture of the desired isomer and its undesired isomer.

After cooling overnight at room temperature, the resulting slurry was dissolved in 100ml of ether: dichloromethane in a ratio of 4: 1 and washed successively with water (50 ml. times.3), 0.2M aqueous NaOH (50ml), water (50 ml. times.2) and saturated brine (50 ml. times.3). After separation of layers, the remaining waste liquid layer (rag layer) was dissolved in dichloromethane. By MgSO₄The combined organic layers were dried, filtered and concentrated on a rotary evaporator at 40 ℃ to give 2.2g of an orange solid. Gas chromatography showed a ratio of desired dialdehyde to undesired isomer of 93: 7. The solid was slurried in hexane and suction filtered to provide 2.15gAn orange solid. This solid was recrystallized from 20ml of refluxing ethyl acetate by cooling to 30-40 c and then cooling in a freezer for 1 hour. The slurry was suction filtered and washed with refrigerator frozen ethyl acetate to give 1.65g of yellow orange needles. mp is 158-160 ℃. The literature mp is 161-162 ℃. Gas chromatography showed a ratio of desired dialdehyde to undesired isomer of 96: 4. Proton nuclear magnetic resonance and carbon nuclear magnetic resonance are consistent with the desired dialdehyde isomer phase.

The yield was 66%.

Preparation of methyl tiglate by thionyl chloride in methanol in proportion and amplification

A mechanically stirred solution of tiglic acid (397.35 g; 3.97mol) treated with pure thionyl chloride (397 ml; 5.44mol) in 3,000ml of methanol was added dropwise over a period of 130 minutes without external cooling, the temperature rising from 14 ℃ to 50 ℃ after 80 minutes. Gas chromatography of an aliquot showed complete conversion to the ester, with no detectable tiglic acid. After stirring for 1 hour at ambient temperature, the solution was distilled under atmospheric conditions through a silvered Vigreux column (400 mm. times.20 mm) with vacuum sandwich. 630ml of condensate which is mainly at 57-61 ℃ is collected when the tank temperature is 58-63 ℃ within 2 hours. Gas chromatography showed a significant amount of methyl esters present in the distillate.

The Vigreux column was exchanged with a less efficient column (30 cm. times.2 cm. w/groove less) to accelerate the distillation speed. 1,300ml of distillate with the head temperature of 65-69 ℃ is collected within 2.25 hours at the tank temperature of 69-71 ℃. Gas chromatography showed significant methyl esters present in the distillate. Atmospheric distillation was continued until the pot temperature reached 87 ℃ during which 975ml of distillate with a head temperature of 69 ℃ to 83 ℃ was collected over 2 hours. Gas chromatography showed that a significantly greater amount of methyl esters were present in the distillate compared to the earlier fractions.

The yellow two-phase mixture in the pot was extracted with ether (300ml and 200ml) by passage K₂CO₃Drying, filtration and concentration on a rotary evaporator at 25 ℃ gave 132.6g of an orange oil (yield 29.3%). Gas chromatography showed the product. Proton nmr and carbon nmr were consistent with the desired product with trace amounts of ether. Gas chromatography of the ether condensate showed some methyl esters present in the overhead fraction.

Distillate 3: the 3 rd methanol distillate (975ml) was concentrated on a rotary evaporator at 25 ℃ to give a two-phase mixture (100 ml-150 ml). The mixture was extracted with ether (100ml and 50ml) by K₂CO₃Drying is carried out.

Distillate 2: the 2 nd methanol distillate (1,300ml) was concentrated on a rotary evaporator at 25 ℃ to give a two-phase mixture (30 ml-50 ml). The mixture was extracted with ether (2X 50ml) by K₂CO₃Drying is carried out.

The concentrated ether extracts of distillate 2 and distillate 3 were combined, suction filtered and concentrated on a rotary evaporator at 25 ℃ to give 77.3g of a colourless oil.

Proton nuclear magnetic resonance and carbon nuclear magnetic resonance were matched to previous spectra of the desired methyl ester.

The total yield was 132.6+ 77.3-209.9 g (46.3%).

Alternatively, 1) methyl tiglate is commercially available from Alfa, Lancaster or Acros. And 2) experiments can be carried out according to JOC, 64, 8051-8053(1999) to prepare the phosphonium salts.

Bromination of methyl tiglate

Heating back with a 1L ball receiver between a 5L reaction flask and a reflux condenserA mechanically stirred mixture of methyl tiglate (209.9 g; 1.84mol), N-bromosuccinimide (327.5 g; 1.84mol) and 70% benzoyl peroxide (3.2 g; 0.009mol) in 2,000ml of carbon tetrachloride was flowed (78 ℃ C. -81 ℃ C.). After 2 hours, the reflux was stopped, the heating mantle was removed and the stirrer was switched off. All solids float in CCl₄On solution, this indicates that succinimide carries negligible NBS. The slurry was cooled to 20 ℃ in an ice bath and suction filtered to afford 180.7g of a milky white solid. No washing was performed. The yellow filtrate was washed with water (1 L.times.3) over MgSO₄Drying is carried out. Gas chromatography showed the starting material methyl tiglate and two monobromides in a ratio of about 1: 2: 1 with other minor components.

Filter off MgSO₄Thereafter, the pale yellow filtrate was concentrated on a rotary evaporator at 35 ℃ to give 327.1g of a pale yellow oil. Proton nmr and gas chromatography showed the following composition:

components	NMR(mole％)	GC (area%)
			Gamma-bromo	50％	49％
Alpha-bromo	26％	21％
			α, γ -dibromo ester (	7％	4％
Tiglic acid methyl ester	6％	10％
			Others	11％	-

The yield of the desired product was adjusted to 46.0% for a 50% analysis.

The use of this oil is the same as in the next step.

The gamma-methyl bromotiglate and triphenylphosphine are subjected to proportional amplification reaction in acetonitrile by using slightly higher effective load

A crude mixture of methyl gamma-bromotiglate (322.6 g; 85% allylic bromide; 1.42mol) in 1,300ml of anhydrous acetonitrile was mechanically stirred in a 5L four-necked flask under nitrogen.

A solution of 2,000ml of triphenylphosphine (387.0 g; 1.48mol) in ethyl acetate was added dropwise over a period of 4 hours. During the addition, the temperature rose from 22 ℃ to a maximum temperature of 30 ℃ after the first 75 minutes of addition of about 40%. After addition of about 60% triphenylphosphine solution during 120 minutes, the solution became cloudy and continued to precipitate a solid out on subsequent additions. After addition, the funnel was rinsed with ethyl acetate (600ml) and added further to the reaction mixture. The milky slurry was stirred at ambient temperature over the weekend.

The white slurry was suction filtered and the filter cake washed with 2: 1 ethyl acetate: acetonitrile (150 ml. times.3). The white solid (352.55g) was dried in a vacuum oven at 40 ℃ for 4 hours (constant weight after 2 hours) to give 322.55g of product. mp 187-188 deg.C (decomposition). Document mp 183 ℃ (decomposition). Proton nuclear magnetic resonance and carbon nuclear magnetic resonance are consistent with previous spectra of the desired phosphonium salt. LC-MS analysis showed a major component whose electrospray mass spectrum in positive ion mode (positive mode) was consistent with the desired phosphonium salt, providing a molecular ion peak at 375 m/z. Phosphorus nmr showed a monophosphoric signal at 22.0 ppm.

The yield based on starting material tiglic acid methyl ester was 100 × 322.55/(455.32 × 1.84 × 322.6/327.1) ═ 39.0%.

Preparation of (3-carbomethoxy-2-Z-buten-1-ylidene) triphenylphosphine

A small, mechanically stirred slurry of (3-carbomethoxy-2-E-buten-1-ylidene) triphenylphosphine onium bromide (154.8 g; 0.34mol) in 3,400ml of deionized water was treated dropwise with a solution of sodium hydroxide (13.6 g; 0.34mol) in 500ml of water at 23 ℃ over a period of 32 minutes, with no significant exotherm, but a pale yellow solid precipitated immediately. After stirring for 15 minutes, the pale yellow slurry was suction filtered, washed with water (1,500ml) and suction dried to give 151.7g of a pale yellow solid. This solid was dried overnight in a vacuum oven at 35 deg.C to 45 deg.C (3:50 pm).

After drying in a vacuum oven at 35-45 ℃ for 22.5 hours, a constant weight of 107.8g is reached. mp 144-160 deg.C, and mp 145-165 deg.C. Proton nuclear magnetic resonance is similar to previous spectra of the desired ylide, taking into account differences in nuclear magnetic resonance field strength. Carbon nmr showed methyl carbons with complex aromatic regions at 50.2ppm and 11.8ppm, with no significant signal for olefinic carbons and ylide carbons.

The yield was 84.7%.

Experimental preparation of dimethyl crocinate

A mechanically stirred mixture of (3-carbomethoxy-2-Z-buten-1-ylidene) triphenylphosphine (12.8 g; 34.2mmol) and 2, 7-dimethyl-2, 4, 6-octatrienedial (2.1 g; 12.8mmol) in benzene (128ml) was heated to reflux using a timer under a nitrogen atmosphere for 6 hours.

The resulting slurry was cooled in an ice bath for 40 minutes, suction filtered, washed with benzene and suction dried to melt the frozen benzene to give 2.1g of a red solid. TLC analysis showed a single yellow spot. The solid was dried in a vacuum oven at 40 deg.C to 45 deg.C for 70 minutes to yield 1.85g of product (40.5% yield). Uncorrected mp is 210-213 ℃. Literature reference³mp-214-216 ℃. (Note:³E.Buchta &andree, Chem be, 93, 1349 (1960)). Proton nuclear magnetic resonance is similar to the spectrum of crocetin on a 90MHz instrument. Carbon nmr showed all 11 unique carbon signals with the correct chemical shift of the desired dimethyl ester and a minor impurity signal, which may be residual benzene. Electrospray mass spectrometry showed fragmentation and recombination of fragments.

TLC analysis indicated that the red filtrate contained additional product triphenylphosphine oxide and one R_fSlightly below the orange component of the isolated solid. The red filtrate was concentrated on a rotary evaporator at 35 ℃ to give 13.2g of a red solid. The solid was heated to reflux in methanol (25 ml). The resulting slurry was then cooled in an ice bath, filtered with suction after 60 minutes and washed with methanol to give 0.6g of a red solid. Drying the solid in a vacuum oven at 45 ℃ for 135 min0.5g of product is obtained. mp is 203-208 ℃. Proton nmr showed the desired diester with residual impurities. Carbon nmr showed only signal of the desired product. TLC analysis showed heterogeneous product spots.

The filtrate was concentrated and stored.

Secondary preparation of dimethyl ester of crocetin

2, 7-dimethyl-2, 4, 6-octatrienedial (11.95 g; 12.8mmol) was added as one portion to a mechanically stirred slurry of (3-carbomethoxy-2-Z-buten-1-ylidene) triphenylphosphorane (73.0 g; 195.0mmol) dissolved in 400ml benzene under a nitrogen atmosphere, followed by an additional 330ml of benzene. The resulting brown slurry was heated to reflux with a timer for 6 hours and cooled to room temperature overnight under a nitrogen atmosphere.

The resulting slurry was cooled to 6 ℃ to 10 ℃ in an ice bath, suction filtered and washed with benzene (50 ml. times.2) to give 10.05g of a red solid. TLC analysis showed a single yellow spot. The solid was dried in a vacuum oven at 40 deg.C (9:00am) for 3.5 hours without weight loss to give 10.05g of product (38.7% yield). mp is 211-214 ℃. Mp is 214-216 deg.C. Proton nuclear magnetic resonance and carbon nuclear magnetic resonance were consistent with previous spectra of the dimethyl ester of crocetin.

The red filtrate was concentrated on a rotary evaporator at 40 ℃ to give 84.4g of a red solid. TLC analysis and experimental work were similar. The solid slurry was liquefied by stirring with magnetic force in 165ml of refluxing methanol. The resulting slurry was then cooled in an ice bath for 2.5 hours, filtered with suction and washed with a minimum amount of methanol to give 10.5g of an orange paste. TLC analysis showed a single yellow spot. The paste was dried in a vacuum oven at 45 ℃ for 190 minutes to give 5.6g of product. mp is 201-208 deg.C. Nuclear magnetic resonance showed the desired diester with unknown aromatic impurities.

This impure solid and a total of 6.5g of two other similar solids from earlier experiments were dissolved in refluxing chloroform (75ml), diluted with methanol and cooled in a refrigerator overnight.

The slurry was suction filtered and washed with a minimum amount of methanol to give 6.1g of a red crystalline solid. The solid was dried in a vacuum oven at 45 ℃ for 3 hours until a constant weight of 4.25g was reached. mp is 211-213 ℃. Proton nuclear magnetic resonance and carbon nuclear magnetic resonance show other olefinic or aromatic impurities. The solid was dissolved in refluxing toluene (150ml) and finally cooled in a refrigerator for 130 minutes. The slurry was suction filtered and washed with toluene to give 2.05g of a red solid. The solid was dried in a vacuum oven at 45 ℃ for 50 minutes without weight change to give 2.05g of product. mp-214-216 ℃. Proton nmr showed the desired dimethyl crocylate with some residual toluene and negligible undesired isomer impurities. Carbon nmr showed the desired dimethyl crocylate without detectable impurities of the undesired isomer and 2-3 new residual signals consistent with toluene. The yield was 45.5%.

Preparation of crocetin disodium salt

A mechanically stirred slurry of dimethyl crocinate (13.95 g; 39.1mmol), 40 wt% aqueous sodium hydroxide (273 ml; 3.915mol) and methanol (391ml) was heated at reflux for 12 hours at 74 ℃ using a timer.

The orange slurry was suction filtered through a cloth funnel with filter paper and a sintered glass funnel. Slow filtration⁴(Note:⁴filtration through sintered glass is fast until the filter is clogged after draining. However, the water wash leaves the filter unobstructed). Adding the slurry in the sintered glass funnel to a distributed funnelOn the solid in (1). The orange paste was washed with water (100 ml. times.3) and then with methanol (50 ml. times.3). The orange paste was dried in a vacuum oven at 45-50 ℃.

After 21 hours, the orange clot weighed 24.25 g. This material was pulverized with a spatula into powder and dried in a vacuum oven at 45-50 ℃.

After 65.5 hours of co-drying, the amount of orange powder was 23.1 g. The infrared spectrum shows an additional band relative to that of the reported TSC, especially at 3424cm^-1And 1444cm^-1Large bands are shown. Proton nmr showed no evidence of methyl esters. However, the integration of the olefinic and methyl zones is separated, which may be due to phase adjustment problems.

The orange solid was magnetically stirred in 400ml of deionized water for 35 minutes, assuming excess weight due to sodium hydroxide. The slurry was suction filtered and the filter cake was washed with deionized water (50 ml. times.2). To provide an orange paste. The mass was dried in a vacuum oven at 45-50 ℃ until constant weight. After about 7 hours, the solid was crushed and pulverized into a powder and further dried in a vacuum oven at 45 ℃ overnight.

After drying at 45 ℃ for 21 hours, the solid content was 13.25 g. After further pulverization into powder and drying at 45 ℃ in a vacuum oven, the solid amount was 13.15 g. The infrared spectrum of the sample is consistent with the reported infrared spectrum. Proton nmr spectra were consistent with disodium salt. HPLC analysis shows a major component that may have a minor impurity. The electrospray anion mass spectrum of the main component is consistent with the required crocetin disodium salt. Carbon nmr showed all 10 unique carbon signals of crocetin disodium salt, which confirmed the symmetry of the molecule.

The initial filtrate of water, sodium hydroxide and methanol precipitated more solids during the water wash. The slurry was suction filtered and washed with water to give an orange paste. The paste was dried in a vacuum oven at 45 ℃ for 18.5 hours to give 0.65g of an orange solid. The spectral data was consistent with the desired disodium crocetinate salt. The solids are combined with the first harvest.

Yield 13.15+ 0.65-13.8 g (94.8%).

Elemental analysis of the first harvest showed that the value of the desired product was unacceptable, indicating that the disodium crocetinate salt was contaminated with sodium hydroxide.

Water washing of crocetin disodium salt

Crocetin disodium salt (13.6g) was slurried in 150ml deionized water and magnetically stirred at room temperature for 1 hour. The slurry was suction filtered onto a Buchner funnel. The orange paste was then washed with water and the pH of the orange filtrate was monitored.

The orange paste was blotted dry on the filter with a rubber bung. The paste was dried in vacuo at 25 ℃ to 55 ℃ for 5.5 hours to give 11.2g of a fluffy orange solid. The solid was pulverized into a powder, transferred to a bottle, and dried in a vacuum oven at 45 ℃ overnight.

The obtained amount was 11.1 g. The recovery rate was 81.6%. The infrared spectrum and proton nuclear magnetic resonance spectrum of the sodium crocetin compound accord with the infrared spectrum and proton nuclear magnetic resonance spectrum of the prior disodium crocetin compound. HPLC analysis showed a single component at 420nm, which is consistent with crocetin in its electrospray mass spectrum in negative ion mode.

Carbon nmr showed all 10 unique carbon signals with the correct chemical shift of the desired crocetin disodium salt. Elemental analysis gave acceptable data for the desired product.

Reference to the literature

1.Tetrahedron Letters，27，4983-4986(1986)；

F.J.H.M.Jansen, M.Kwestro, D.Schmitt & J.Lugtenburg, Recl.Trav.chem.Pays-Bas, 113, 552-562(1994) and references cited therein;

3.j.h.babler, US patent No.4,107,030, Apr 21, 1992;

4.T.W.Gibson & P.Strassburger，J.Org.Chem.，41，791(1976)& J.M.Snyder & C.R.Scholfield，J.Am.Oil Chem.Soc.，59，469(1982)。

example 6

Purity characterization of TSC prepared according to the improved Synthesis method

For the TSC material synthesized according to the method of example 5, the ratio of absorbance at 421nm to absorbance at 254nm, as determined using an uv-vis spectrophotometer, was 11.1.

Example 7

Oral administration of TSC

When TSC was administered orally (via gavage techniques) in rats, it was shown that TSC was absorbed into the bloodstream. In both rats, 1% to 2% of the administered dose was found to be present in the bloodstream from 15 minutes to 30 minutes after administration. The maximum amount of oral absorption actually occurs before this time.

It will be apparent to those skilled in the art that numerous modifications and additions can be made to the compounds and compositions of the present invention and related methods without departing from the disclosure thereof.

Claims (30)

1. A method for converting an isomeric mixture of an enedialdehyde to an all-trans aldehyde comprising isomerizing the isomeric mixture of dialdehydes with sulfinic acid in a solvent.

2. The method of claim 1, wherein the sulfinic acid has the formula RSO₂H, wherein R is a linear or branched alkyl or aryl group of C1 to C10.

3. The process of claim 1, wherein the solvent is selected from 1, 4-dioxane, tetrahydrofuran, or dialkyl ethers wherein the alkyl group is a linear or branched alkyl group of C1 to C10.

4. The process of claim 1, wherein the sulfinic acid is p-toluenesulfinic acid and the solvent is 1, 4-dioxane.

5. The method of claim 1 wherein the enedialdehyde is 2, 7-dimethyl-2, 4, 6-octatrienedial.

6. The process of claim 1, wherein the olefinic dialdehyde is 2, 7-dimethyl-2, 4, 6-octatrienedial, the sulfinic acid is p-toluene sulfinic acid and the solvent is 1, 4-dioxane.

7. A method of synthesizing a bipolar trans carotenoid salt compound having the formula

YZ-TCRO-ZY，

Wherein:

y is a cation, and Y is a cation,

z ═ a polar group bound to the cation, and

TCRO ═ trans carotenoid skeleton,

the method comprises the following steps:

a) coupling symmetric dialdehyde containing conjugated carbon-carbon double bond with triphenyl phosphorane,

b) saponifying the product of step a).

8. The process of claim 7, wherein the coupling of step a) is performed using [ 3-carbomethoxy-2-buten-1-ylidene ] triphenylphosphorane.

9. The process of claim 7, wherein the product of step a) is saponified with a solution of NaOH and methanol.

10. The process of claim 7 wherein step a) is followed by a step of isolating the desired product of the coupling reaction.

11. A process for the saponification of a symmetrical diester containing conjugated carbon-carbon double bonds to form a bipolar trans carotenoid salt compound, the process comprising the steps of:

a) dissolving the symmetrical diester containing conjugated carbon-carbon double bonds with a compound selected from the group consisting of methanol, ethanol, propanol and isopropanol, and

b) mixing the solution of step a) with a base.

12. The method of claim 11, wherein the base is selected from the group consisting of NaOH, KOH, and LiOH.

13. The method of claim 11, wherein the diester is saponified with methanol and NaOH.

14. A bipolar trans carotenoid salt compound synthesized according to the method of claim 7.

15. A bipolar trans carotenoid salt compound according to claim 14 which is trans sodium crocetinate.

16. A trans sodium crocetinate composition wherein the absorbance of the highest peak occurring in the visible wavelength range divided by the absorbance of the peak occurring in the ultraviolet wavelength range is greater than 7.5.

17. An inhaler comprising trans-sodium crocetinate.

18. A method of solubilizing a bipolar trans carotenoid salt compound, said bipolar trans carotenoid salt compound having the structure:

YZ-TCRO-ZY，

wherein:

y is a cation, and Y is a cation,

z ═ a polar group bound to the cation, and

TCRO ═ trans carotenoid skeleton,

the method comprises the following steps:

a) a dilute solution of sodium carbonate or sodium bicarbonate is prepared,

b) the dilute solution is added to deionized water to raise the pH to 7 or above 7,

c) adding the bipolar trans carotenoid salt compound to the solution of step b).

19. A method of solubilizing a bipolar trans carotenoid salt compound, said bipolar trans carotenoid salt compound having the structure:

YZ-TCRO-ZY，

wherein:

y is a cation, and Y is a cation,

z ═ a polar group bound to the cation, and

TCRO ═ trans carotenoid skeleton,

the method comprises the following steps:

a) adding the bipolar trans carotenoid salt compound to a salt solution,

b) undissolved material was removed.

20. A method of solubilizing a bipolar trans carotenoid salt compound, said bipolar trans carotenoid salt compound having the structure:

YZ-TCRO-ZY，

wherein:

y is a cation, and Y is a cation,

z ═ a polar group bound to the cation, and

TCRO ═ trans carotenoid skeleton,

the method comprises the following steps:

a) a base is added to water to prepare an alkaline solution,

b) adding the bipolar trans carotenoid salt compound to the solution.

21. A method of solubilizing a bipolar trans carotenoid salt compound, said bipolar trans carotenoid salt compound having the structure:

YZ-TCRO-ZY，

wherein:

y is a cation, and Y is a cation,

z ═ a polar group bound to the cation, and

TCRO ═ trans carotenoid skeleton,

the method comprises the following steps:

a) preparing the deionized water, namely preparing the deionized water,

b) adding the bipolar trans carotenoid salt compound to the solution of step a).

22. The method of claim 18, 19, 20, or 21, wherein the compound is trans sodium crocetinate.

23. Use of a bipolar trans carotenoid salt compound in the manufacture of a medicament selected from the group consisting of medicaments for: increasing oxygen diffusivity in a mammal, treating a respiratory disease, treating emphysema, treating hemorrhagic shock, treating cardiovascular disease, treating atherosclerosis, treating asthma, treating spinal cord injury, treating cerebral edema, treating papilloma, treating hypoxia, treating ischemia, treating traumatic brain injury, improving function in a mammal, treating diabetes, or treating Alzheimer's disease.

24. Use according to claim 23, wherein the medicament for use in increasing the diffusivity of oxygen in a mammal is administered by inhalation.

25. The use as in claim 23 wherein the bipolar trans carotenoid salt compound is trans sodium crocetinate.

26. Use of a bipolar trans carotenoid salt compound in the manufacture of a medicament selected from the group consisting of medicaments for: increasing diffusivity of oxygen in a mammal, treating emphysema, treating hemorrhagic shock, treating ischemia in a mammal, treating traumatic brain injury, improving function, treating diabetes, or treating alzheimer's disease, wherein absorbance of the highest peak occurring in the visible wavelength range divided by the absorbance of the peak occurring in the ultraviolet wavelength range of the bipolar trans carotenoid salt compound is greater than 7.5.

27. The use of claim 26 wherein the bipolar trans carotenoid salt compound is trans sodium crocetinate.

28. Use of a bipolar trans carotenoid salt compound in the manufacture of a medicament for treating, preventing or reducing the amount of ischemia caused by surgery in a mammal, wherein a therapeutically effective amount of the bipolar trans carotenoid salt compound is administered to the mammal before, during or after surgery.

29. The use of claim 28 wherein the bipolar trans carotenoid salt compound is trans sodium crocetinate.

30. Use of trans sodium crocetinate in the manufacture of a medicament for increasing the diffusivity of oxygen in a mammal, wherein the medicament is administered to the mammal by inhalation a therapeutically effective amount of trans sodium crocetinate.