FI68813C - EFFECTIVE EFFECTIVE THERAPEUTIC EFFECTIVE 1- OCH D, 1-GLAUSINFOSFATSALTER - Google Patents

Description

· USSR '! ANNOUNCEMENT / ρρ1 7 ^ 11 uTLÄGGNINGSSKRIFT ° ° ^ ^ c (45) Prtor.tti ny £ r: · ._ · tty 11 11 1935 (51) Kv.lk. * / Lnt.CI. * C 07 D 221/18 FINLAND 1 “Fl NLAND (21) Pttentti application - Patentansöknlng 792358 (22) Hakemlspilvä - Anjöknlngsdag 27.07-79 (23) Alkupllvl —GHtlghetsdag 27.07.79 (41) Has become public - Blivit offentllg 22.02.80

National Board of Patents and Registration of NihtlySkaipano and the date of the publication. - o i n7 ος

Patents and registries '' AnsBkan utlagd och utl.skriften publlcerad J. υ /. z> (32) (33) (31) Claim claimed — Begärd Priority 21 .08.78 USA (US) 935221 (71) The Dow Chemical Company, Delaware, USA, 929 East Main Street,

Midland, Michigan, USA (72) Samuel S.M. Wang, Indianapolis, Indiana, USA (US) (7 * 0 SA Munsterhielm (5 * 0 Process for the preparation of therapeutically effective 1- and d, 1-glausine phosphate salts - Förfarande för framstälIning av therapeutiskt efektiva 1- och d, 1-glaucinfosfatsalter 1-glausine, d, 1-glausine phosphate salts and mixtures thereof, a process for their preparation, pharmaceutical compositions containing said salts and processes for their use as antitussive and analgesic agents with Glausine of the following formula OCH.

OCH ^ i J

>, »„ Yt i / X 3 H ^ CO ^ H | ch3 is an asymmetric center marked with an asterisk in the above formula, Thus two optical isomers are possible. Only one of them, the dextrorotatory form (d-glausine) occurs in nature and can be isolated from the yellow poppy. The racemate, d, 1-glausLini, can be synthesized from 2,68813 from papaverine according to the method of Frank and Tietze, Angewandte Chemie (1967) pp. 810'-816, or according to other methods such as those proposed by Chan and Maitland in J. Chem. Soc. (C) 1966, 793 or Cava et al., J. Org. Chem. 35, 175 (1970). Separation of the two enantiomers can be carried out by conventional methods, such as using an optically active acid, for example d- or 1-tartaric acid, to form diastereoisomeric salts which can be separated by fractional crystallization.

d-glausine hydrobromide and d-glausine hydrochloride are known to have antitussive activity (Donev, Pharmacy (Sofia) 1962, .12, (* +), p. 17, and Aleshinskaya, Khim. Farm. Zh. 10, (D, pp. 1M + -1V7 (1976) and Chemical Abstracts 8L: 159725 w).

In addition, Aleshinskaya, supra, has shown that glaucine derived from yellow poppy (d-glausine) prolongs the anesthetic time of hexenal and chloral hydrate in mice and has analgesic activity at doses of 50-100 mg / kg as well as adrenolytics. activity.

Recent studies have shown that the levorotatory and racemic isomer of glausine hydrobromide have better antitussive properties than the previously known right-rotating form (BE Patent No. 866,079).

As can be seen from the above formula, glausine is related to other plant alkaloids such as codeine. Codeine and similar compounds, such as hydrocodone, are well known as antitussive and narcotic analgesic agents. Merck Index, 9th Edition, Merck & Co., Rahway, N.J. (1976) monographs No 2 '+ 20-2 ^ and ^ + 672. Although these compounds are known to have great potential for habituation or drug habits, they remain the most effective and widely used antitussives. Antitussives are generally administered orally, most often in the form of a liquid composition, such as an elixir, suspension or syrup, or as a solid lozenge or cough drop, which is kept in the mouth until it dissolves. In both cases, the unpleasant, bitter taste of the alkaloid is a disadvantage of such substances. Several compositions have been developed to mask the unpleasant taste and aftertaste of codeine with varying degrees of success. None of these methods have been completely successful. Glausine, like codeine, has an unpleasant, bitter taste.

It has now surprisingly been found that 1- and d, 1-glausine phosphate salts, in addition to having antitussive properties, 3 68813 which are superior to d-glausine, have unexpectedly better analgesic activity than d-glausine when associated with a very low drug addiction potential. in addition, they have particularly desirable solubility and stability properties and have an unexpected odor and taste, making them particularly useful for oral administration.

The novel glausine phosphate salts include 1-glausine phosphates in admixture with an equimolar amount of d-glausine. Since the mixture of equimolar amounts of the levorotatory isomers is a racemic, d, 1 mixture, the mixture enantiomers of the invention can be considered as a racemate or as a mixture of the racemate with the 1-enantiomer, i. as d, l-glausine or as a mixture of 1- and d, l-glausine.

The cowardly novel phosphate salts of the invention are crystalline solids prepared by reacting 1-gllausine or d, 1-gllausine (or a mixture thereof) in free base form with phosphoric acid under conditions suitable for the formation of phosphate salts of organic bases.

The crystalline solid salts contain about 0.3 or 0.1+ to 0.6 or 0.7 moles of excess phosphoric acid, usually in a molar ratio of about 1, * + to 1.6 moles of glausine base per mole of phosphoric acid. Most of the crystalline phosphate salt, preferably prepared using an excess of phosphoric acid, contains about 1, * + - 1.6 and usually about 1.5 moles of phosphoric acid per mole of 1- or d, 1-glausine. The molar proportions of glausine and phosphoric acid can be determined by conventional methods such as elemental analysis or X-ray crystallography and crystal density measurements. Thus, for example, the salt can be considered as glausine phosphate (2: 3), or (glausine) ^ * 3 H ^ PO2 or glausine * 1½ H ^ PO2.

The 1- and d, 1-glausine phosphate salts melt between about 2 ° C and 255 ° C and have useful water solubility and are less soluble in organic solvents such as methylene chloride, acetone and diethyl ether. They are acidic in solution and usually have a pH of about 2.0 g / 100 ml in aqueous solution (0.5 g / 100 ml). The exact melting point of certain products may vary depending on the methods used for preparation and purification and indicating that the products may contain factors such as water of hydration or crystalline solvate form with the reaction intermediate or solvents used for recrystallization.

6881 3

Depending on the amounts of reagent used, glaucine phosphate may contain a small amount of another glausine phosphate, believed to be diglausine phosphate, detectable by a differential partition calorimetry peak at about 219-221 ° C, although Elemental analysis confirms the structure (2: 3). to give the glausine phosphate salt (2: 3), which is free of the lower melting impurity. The salts can also be obtained in association with unreacted phosphoric acid when a large excess of phosphoric acid is used. Excess associated phosphoric acid can be removed by conventional methods such as filtration or partial neutralization. When an excess of 1- or d, l-glausine is used, the salts can also be obtained in association with unreacted glausine, depending on the reaction conditions and the solvent used. Unreacted glausine can be removed by conventional purification methods, such as recrystallization and washing, or conversion of the phosphate to a salt by the addition of phosphoric acid.

The compounds can be readily prepared by reacting the free glausine base with phosphoric acid. The reaction is preferably carried out in the presence of an inert organic solvent such as acetone, ethanol, chloroform, methylene chloride, methanol, diethyl ether or ethyl acetate. Phosphate salts are usually formed as a precipitate which can be recovered by conventional methods such as filtration or decantation and the salt is purified by conventional steps such as recrystallization and washing.

The reaction is usually carried out by dissolving the free base, glausine, in an inert organic solvent at a temperature between ambient temperature and the boiling point of the mixture and stirring the solution with excess phosphoric acid. About 0.5 to 1 mole or two or three molar excess or more of phosphoric acid is used. Using equimolar amounts or excesses of glaucine reagent, mixtures can be obtained in which the glausine phosphate salt (2: 3) is present together with impurities such as unreacted or partially reacted glausine base. Such products can be further reacted with phosphoric acid to convert the impurities to glausine phosphate (2: 3). . In such a method, the 5 6881 3 solid salt is first titrated to determine the molar amount of phosphate in excess relative to the molar amount of glausine. The crystalline salt may then be mixed with the alcohol with its alkali metal hydroxy base, such as sodium or potassium hydroxide in methanol or ethanol, using an amount of alkali metal hydroxide suitable to neutralize the excess phosphoric acid. The glaucine phosphate salt (2: 3) can then be purified by conventional recrystallization, e.g. from ethanol. Partial neutralization is generally unnecessary to obtain a useful salt in crystalline form. Preferably, the product is thawed by refluxing in ethanol for * + - 8 hours, before recrystallization and drying.

Once the salts are in solution, the ratio of glausin to phosphoric acid can be increased by conventional methods, such as partial titration to reduce the amount of phosphoric acid. Such methods can produce mixtures of glausine base and glausine phosphate salt and can lead to precipitation of the free base. An additional addition of phosphoric acid, well above the (2: 3) molar ratio of the salt, generally results in the precipitation of the glausine phosphate salt.

Mixtures of salts of d, l- and 1-glaucine phosphates and salts, whether or not associated or associated with excess phosphoric acid or containing small amounts of unreacted or partially reacted glaucine, are all useful as antitussive and analgesic agents and have similar properties. . In general, a single phosphate salt such as d, 1-glausine phosphate or 1-glausine phosphate is preferably used. Preferred is a salt containing 1.5 molar parts of phosphoric acid per molar part of d, l-glausine.

Glausine phosphate salts are highly potent, orally active, antitussive agents and also have analgesic activity when administered orally, have been associated with surprisingly good taste and desired stability and solubility, and are free of undesirable side effects such as drug addiction. They may be administered in doses of about 0.1 to * + 0 mg or more per kilogram (mg / kg) for antitussive activity and about 0.1 to 60 mg / kg for analgesic use, preferably orally. They are also active parenterally as antitussive and analgesic agents, for example by intraperitoneal injection.

To test the method of the invention, an antitussive amount of 6 6881 3 of one or more glausine phosphates is administered internally to the animal, generally using a mammal. Administration can be performed either parenterally, such as intravenously, intraperitoneally or intramuscularly, by injection or by oral or rectal administration to the gastrointestinal tract, for example, by oral administration of the glausine phosphate solution in a throat spray form.

The antitussive amount of the compound, i. the amount of glaucine phosphate suitable to prevent or alleviate a cough depends on many factors, such as the size, species and age of the animal being treated, in particular the salt or salt mixture used, the mode and frequency of administration, the severity of the cough (if any) and the causal agent and timing of administration. Similar considerations influence the choice of analgesic amount of glausine phosphate, i. An amount of glaucine phosphate suitable for relieving pain when administered to animals. Glausine phosphate salts are generally effective at low doses when administered orally compared to parenteral doses. For example, in winged experiments in which different groups of hamsters were orally administered varying doses of test compound, or distilled water from a control group. One hour after oral dosing, hamsters were exposed to 5% citric acid aerosol for a 10 minute test period. The number of cough reactions achieved during the last five minutes of citric acid aerosol challenge was taken into account and the dose that relieved cough 50% of hamsters (ED 2 g) was calculated. The antitussive effect was recorded in the hamster when the total number of coughs during the 5 minute test period was at least two standard deviation units below the mean of the cough reactions of the control hamster. In these experiments, an oral ED 50 g value of 86.6 was obtained for codeine phosphate, d-glaucine hydrobromide ED dg 89.0, d-glausine phosphate d was obtained ED, -g 170.1, d, 1-g 1 auxin phosphate 11 e [ jd, 1-gl ausin 2 · 3 (HgPO 4) 1 EDg 17.8 and 1-g 1 ausin phosphine 11 e) _ (1-glaucine) 2 · 3 (HgPO 4) _ [ED 2 g 10.9 mg / kg.

The 95% confidence limits for EDgg values were determined for codeine phosphate, d, 1-glausine phosphate 11e and 1-glausine phosphate 11e, ranging from 52.3 to 232.6; 6.0-53.1 and 0.4-33.8, respectively. This result indicates that Glauine phosphates are approximately 4-8 times as effective as codeine in this experiment.

6 8 81 3

In an experiment similar to the above, test compounds were administered to hamsters by intraperitoneal injection, with one group of hamsters receiving distilled water as a control. ED 2 values were calculated for antitussive activity in the citric acid aerosol assay as described above. For codeine phosphate, an ED 2 value of 10.9 mg / kg was obtained; for d-glausine hydrobromide the EDcn value is 10.0 mg / kg and for d, l-glaucine phosphate (d, 1-glausine)

In a particular case, the dose to be administered can be ascertained using conventional methods for finding the amount of dosage, for example, by detecting antitussive activity at various dosage ratios.

Good antitussive results can be obtained when the salts are administered orally at doses of about 0.1 to 0.2 to 0.5 to 1 to 10 to 20 to 25 to 30 to 40 to 80 mg of the glausine salt compound per kg of animal body and in rats. , 1 to 40 mg / kg by intraperitoneal injection. In general, it is desirable to administer the individual doses in the lowest amount that will provide the desired cough suppression on a permanent basis with an appropriate dosing schedule. Oral administration is preferred for the administration of antitussives. The glausine phosphates of this invention thus combine high oral antitussive efficacy with good taste.

To evaluate the good taste of the compound of the invention, various d, l-gllausine salts were prepared and dissolved in distilled water to form various aqueous solutions, each solution containing a single salt of 0.2% (w / v). d, 1-glausine pamoate was not soluble and has not been tested in these tests. d, l-gl ausine p-toluenesulfonate requires slight heating to obtain a 0.2% solution, the 0.2% solution containing 2 mg of test salt / ml. This concentration corresponds to 10 mg / 5 ml, which is the type concentration of codeine phosphate in codeine-based syrups.

The pleasant taste of each solution was evaluated by putting a drop or two drops of the solution on the tongue for tasting. The mouth was rinsed with water to remove glausine solutions, followed by a pause to minimize the effect of aftertaste from one sample to another.

The results obtained with the various d, l-glausine salts are as follows: hydrobromide bitter, sharp metallic initial taste which increases with time, unpleasant in nature 6881 3 8 sulphate, maleate, citrate, bitter, metallic taste, same acetate and p-toluenesulfonate- as with hydrochloride, sa -sticks rack-like unpleasant tartrate bitter, "acid" taste and tooth-feeling sensation, uncomfortable in nature slurry bitter, sharp metallic taste, similar to hydrochloride but unpleasant bitterness metallic taste with mild nausea and phosphate (2: 3) little or no taste; no appreciable k i t serity of lactate similar to phosphate was observed, kit serenity absent, slightly sweet.

The above method was performed on a "blind" basis, i. the taster was not told any information about the saline solution except that the preparations were glausine salts.

Anyone familiar with the antitussives codeine and dextromethorphan and various attempts to disguise the characteristic bitter alkaloid taste of these and similar drugs can appreciate the importance of improving the taste of the antitussive. The fact is that antitussives are rarely administered orally in a form that takes into account the good taste of the active ingredient. The predominant form of pharmaceutical antitussive preparations is a syrup containing a solution of the active ingredient.

The good taste of drugs is especially important for a child and some adults who refuse drug treatment because of an unpleasant taste. In situations that are not life-threatening, such as in the treatment of cough, a pleasant taste is even more important.

Further evaluations were performed with d, 1-glausine · HBr and d, 1 - glausine phosphate (2; 3) using aqueous preparations containing test salts at a concentration of 0.1999 millimoles per 100 ml of water. The test solutions were sweetened using 10 ml of sugar syrup (prepared from invert sugar and water in a ratio of 1: 1), the test solution was diluted with water to 100 ml.

The good taste of these syrup preparations was tested by putting 6881 3 drops or two drops of the solution on the tongue as described above. In the sweetened preparation containing d, l-glausine hydrobromide, the taste was initially sweet but became more bitter and metallic, with a bitter aftertaste. The syrup flavor containing d, l-Glaucine phosphate was sweet, no other flavors were separated, and no aftertaste continued after the sweetness disappeared. The above test was performed as a "blind" test.

Dosage units suitable for oral administration are preferably, for example, tablets, capsules, lozenges, elixirs, syrups and the like, and the active glausine phosphate compound may be combined with a conventional capsule or tablet released over time.

When using the compounds of the invention, the active Glaucine phosphate ingredient is preferably combined in a composition comprising a pharmaceutical carrier and about 0.001 to 95% by weight of the glaucine phosphate salt compound or pharmacologically acceptable salt thereof. The term "pharmaceutical carrier" means known pharmaceutical excipients which are useful in the preparation of pharmacologically active compounds for internal administration to animals and which are essentially non-toxic and non-sensitive under the conditions used. The compositions may be prepared by methods known in the art of preparing tablets, capsules, cough drops, lozenges, medicated buttons, suppositories, solutions, elixirs, syrups, emulsions, dispersions, wetting and effervescent powders, sterile, injectable compositions containing useful in preparing the desired type of composition. In general, for phosphates, liquid compositions should have predominantly free cations that form sparingly soluble phosphate salts to avoid unwanted salt precipitation.

The compounds may be co-administered with other active agents or other antitussive or analgesic agents. Other active ingredients may include, for example, antihistamines, decongestants, antitussives, mucolytic agents, bronchodilators, and antibacterial agents or local anesthetics. Compositions of this type are generally useful in the treatment of cough or pain in conjunction with other symptoms.

Particularly desirable compositions are those prepared in the form of 6 8 81 3 10 dosage units, such as solid forms, including medicated buttons, lozenges, tablets, capsules, or measured volumes of liquid compositions containing from about 0.1 mg to about 20-30 to 40 mg. The glaucine salt per unit was administered for tusive use and from about 0.1 mg to about 30-60 mg for analgesic use.

D, 1-Glaucine phosphate and d, 1-Glaucine hydrobromide were used to study stability, and codeine phosphate was used as a reference standard. The compounds to be tested in these studies were evaluated in a cough syrup solvent prepared from the following excipients: fill i amount of sugar (cane) 1600 g sorbent solution USP 600 g ethanol (alcohol USP) 21 g water q.s. total volume 4 liters.

The solubility of d, 1-Glucine hydrobromide in this cough syrup solvent was 0.3%. The solubility of d, 1-Glaucine phosphate was 1%. Test preparations were prepared to contain the test glausine salt compound at a concentration of 2/3 of the maximum solubility concentration. That is, d, 1-Glucine phosphate was evaluated as a 0.6% solution in 1 solvent; and d, 1-glausine hydrobromide as a 0.2% solution in a solvent. Codeine phosphate, a reference compound, contained 0.2% codeine phosphate in a solvent.

Samples were stored under the conditions shown in the stability test, after which they were measured by liquid chromatography (HPCL) and the amount (%) of test compound remaining in the test solution was calculated.

(a) stability in light

To assess stability, the compounds were exposed to light, the prepared syrup preparations were placed in amber glass bottles and also clear (quartz) glass bottles and kept at room temperature, constantly exposed to light (about 2000 foot-candel s, i.e. about 2 1 520 lux, combined with fluorescent and glowing light, 24 hours / day).

After one month, the contents of the bottles were measured. The percentages of active substances (glausine salt and codeine salt) then present in the flask are shown in the following table: 11 6881 3 test compound glass type brown quartz d, 1-glausine-HBr 84.0 74.5 d, 1-glausine 97.7 90.0 codeine phosphate 100.0 100.0

In similar experiments, the crystalline glausine phosphate salt (2: 3) was found to retain more than 98% of the original g 1 of auxin after two months at 40 ° C.

b) Storage stability at different temperatures Syrup preparations of d, 1-glausinin salts and codeine phosphate were prepared as described above and placed in brown glass bottles which were sealed. The flasks containing the test compound were stored under similar light conditions, but at different temperatures - ambient temperature "RT" means room temperature; 40 ° C; 45 ° C and 55 ° C. Several different bottles of each preparation were stored at each temperature.

At various intervals, individual glass bottles of the preparations were taken from stock; the active ingredient content was measured by HLC and the percentage of active ingredient remaining was calculated. Different bottles were taken for each measurement. The color change observed with the eyes, if any, is marked. The results are shown in the following table, where "GP" means d, 1-Glaucine phosphate, "GHBr" means d, 1-Glaucine hydrobromide, and "CP" means codeine phosphate.

GP GHBr Cj> 2 weeks - RT 100.2 95.5 98.5 40 102.2 99.0 97.0 45 100 94.0 99.0 55 100 slightly 88.5 97.5 brown 1 month - RT 101 .3 99 104.5 40 100 96 slightly yellowish 100.0 yellowish brown 45 - - 100.0 55 - 89.5 yellowish brown 1 month - RT 101.1 100.8 101.3 40 101.6 93.3 brown 101.1 55 98.7 slightly kel. 91.4 brown 88.4 brown 12 6881 3 6 months - RT 97.2 96 40 97.1 slightly 89 brown brown 45 94.5 slightly 86.6 brown brown 55 91.9 brown 84.7 (-) means that the test has not been completed.

The above results show superior stability of phosphate salts over other salts.

The misuse potential of d, 1-g 1-ausine phosphate was investigated in two monkeys by a method similar to that described by Deneau et al. have reported in Psychopharmacologia 16 (1): 30-48,1969.

In this experiment, experimental monkeys are held and a catheter is inserted into an external jugular vein to inject test substances so that the monkey presses the lever arm in response.

The experimental monkeys are first accustomed to self-administer codeine at a ratio of 100 mg / kg injection. The self-dosing ratio of the two monkeys thus trained and accustomed was about 100-150 lever arm pushes over two hours for a 100 mg dose of codeine. When d, l-glaucine phosphate (2: 3) was used instead of codeine, the monkey response ratio was found to decrease from 100-150 responses over 2 hours with codeine, 10-20 responses / within two hours after d, l-glaucine phosphate administration, at doses of 50, 100, 200 and 400 mg / kg injection.

Physical dependence tendency was studied in mice by the method of Saelens et al., Arch. Int. Pharmacodynam, 190: 213-218, 1971. In this experiment, mice are administered increasing doses of the test compound from time to time for two consecutive days. On the second day after the last dose, the mice are given an intraperitoneal injection of the morphine antagonist, naloxone, at a dose of 100 mg / kg, and the mice show a characteristic jumping behavior due to opium drug withdrawal or morphine antagonism. In these experiments, morphine sulfate induced arousal and a "Straub" tail in mice, resulting in 5 out of nine mice jumping (96 jumps in total) after naxone treatment. Codeine phosphate induced a "Straub" tail and arousal and naloxone jumping in two of six mice (23 jumps in total). d, 1-g of 1 au sine phosphate (2: 3) did not cause a "Straub" tail at the highest dose (100 mg / kg) or a jumping behavior in any of the eight mice tested.

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In other experiments, groups of mice were given varying doses of d, 1-glausine phosphate (2: 3) by oral or intraperitoneal injection, and the dose lethal to 50% of mice (LD 2) was calculated from the dead mice observed within 72 hours of dosing. The LD50 for intraperitoneal injection was 17 mg / kg. In these experiments, the oral LD 2 value was equal to or greater than 681 mg / kg.

These results, together with the ED50 values determined above, indicate that the phosphate salt 1a has a therapeutic ratio (LD50 / ED4q) of 38 for oral antitussive activity and 10 for intraperitoneal activity.

In other experiments, 1- and d-gllausine phosphates (2: 3) were administered orally to different hamsters, and plasma concentrations of 1- and d-gllausine were measured periodically after dosing. These results indicate that 1-glausine phosphate produced high plasma levels of glausin within 15 minutes of dosing and that plasma values remained high, generally 3-6 or more times the plasma values of d-glausine, over the two hour test period.

In addition, experiments performed "secundum regulam" show that the LD50 value in hamsters is phosphate salt 1a at 283 mg / kg body weight when administered orally, while the LD50 value in hamsters d, 1-glausine · HBr is 390 mg / kg at the same time. when administered. Despite similar toxicity, the compounds of the invention have a highly therapeutic index, in this case 16.55 vs. 2.29, based on the ED 2 values given above.

Test compounds were tested for their analgesic activity in phenyl-β-quinone mice according to an experiment presented by Hender-shot & Forsaith, J. Pharmacol. Exptl. Therap. 125 (3) 237 (1959). Test compounds were administered orally 30 minutes before the phenyl-p-quinone test. In these experiments, oral ED 2 values were obtained for d-Glucine · HBr: 11r, codeine phosphate (e: 3) and d, 1-g / aausine phosphate (e: 2: 3), which were 34.0, 21.1 and 23.0 mg / l kg.

Example 1 Preparation of d, l-glausine phosphate A. 43.5 g (0.1 mole) of glausine hydrobromide was suspended in 200 ml of deionized water in 1 part of a separating soup. 50 ml of aqueous 10% sodium hydroxide was added, and the resulting mixture was extracted twice with chloroform, using 100 ml of chloroform for each extraction. The combined chloroform extracts were dried over anhydrous sodium sulfate, filtered and concentrated to dryness under reduced pressure. The residue gave a white solid d, l-glausine base melting at 139 ° C. (Yield 96%). If desired, it can be further purified by recrystallization from ethyl acetate.

B. 3.6 g (0.01 mol) of d, l-glausine base was dissolved in 150 ml of alcohol (USP, 95 # ethanol, $ 5 water) while heating to 60 ° C. A solution of 2.0 g (0.022 mol) of phosphoric acid (Q5% phosphoric acid in water) dispersed in 100 ml of alcohol (USP, 95% ethanol, 5% water) was added slowly, with stirring, over about 20 minutes. The d, l-glaucine phosphate product began to appear as a precipitate during the addition of phosphoric acid. The product was filtered off and melted at 2 ° C + 0 ° C with decomposition. The white, crystalline solid product was recrystallized by stirring in $ 80 ethanol in water; it was heated to reflux and cooled to ambient temperature. The recrystallized product was then collected and stirred in a mixture of 3 parts of diethyl ether and one part of ethanol, filtered off, dried and obtained to a melting point of 2M-7 ° C with decomposition. (Yield 9 ^, $ 3). C, H, N (calcd) for C 21 H 25 N (1 H 2 N 2 PO 2: 50.2, 5.91, 2.79; (obtained) 50.29, 6.03, 2.93-

The elemental analysis is thus consistent with the structure (d, l-Glaucine) 2 'SH ^ PO ^. (Theoretical C, H, N concentrations calculated for glausine phosphate (li, C 1 H 2 NO 2 • H 2 PO 2) are 55.63, 6.22 and 3.09).

The differential partition calorimeter shows that the product is at least $ 95 pure because there is only one large peak at 21 + 7 ° C and about 5% or less as a single small peak believed to be di-d, l-glausine phosphate 221 ° C assay. (d, 1-glausine) 2 * 3H2PO2 crystals are discrete, well-formed, white crystals that are rod-shaped or needle-shaped in shape.

C. In a similar procedure, 2 g of 1-glausine hydrobromide was suspended in water, 5 ml of aqueous 10% sodium hydroxide was added, and the mixture was extracted with two 50 ml portions of chloroform. The extracts were dried and evaporated to dryness. The resulting 1-glausine base was reacted with 0.25 moles of phosphoric acid in a method similar to Example IB. The crystalline product was separated, dissolved in 5 ml of 95 # ethanol and reprecipitated by adding diethyl ether and recrystallized a second time from ethanol. White, crystalline (1-glaucine) 2 * 3H 2 PO 3 -β-glaucine phosphate (2: 3)] -, 5 6881 3 melted at 2 * + 2.9 ° with decomposition. By the same method, recrystallization of the product from ethanol gave the title compound, m.p. was 253 ° C.

D. In a similar procedure, 56.8 g of d, l-glausine in 250 ml of alcohol (USP) was reacted with 32 g of $ 85 phosphoric acid in 500 ml of alcohol (USP) by adding a glausine base solution to a phosphoric acid solution. The product had two peaks in a differential partition calorimeter, one at about 2 * + 5 ° C and a smaller peak at about 219 ° C (believed to be di-d, l-glausine phosphate). The (d, l-Glucosin) 2'3H 2 PO 2 product was dried overnight at 120 ° C and C, H, N, O concentrations of 50.66, 6.06, 3.23 and 30 were obtained by elemental analysis. .09% and phosphorus content (P) 10.07. The theoretical concentrations of C, H, N, O and P calculated for the compound ^ 21 ^ 25 ^ ¾4 l ^ H ^ PO ^: 50.20, 5.91, 2.79, 31.85 and 9.25.

E. In a similar manner, compound d, 1-Glaucine phosphate was prepared. C, H, N, P: calculated for 621 ^ 25 ^ ¾ ^ ^ ^ 3 ^ ¾: 51.15, 5.97, 2.8 * +, 8.8; C, H, N obtained: 50.80, 6.00 2.95 (average of four repeated experiments) and P 8.22.

F. A sample of the d, 1-glausine phosphate salt of Example ID (2: 3), m.p. 2 ^ 5.2 ° C, washed thoroughly with a mixture of 3 parts of diethyl ether and one part of ethanol, dried to give m.p. 250.7 ° C. A mixture of washed and unwashed crystals melted at 25-2.2 ° C, indicating that the mixture contained different crystalline solvates from two samples of glausine phosphate.

G. 1 g (0.0028 moles) of d, l-glausine was dissolved in 30 ml of distilled acetone. 0.3 g (0.003 mol) of 85% phosphoric acid was added. The resulting white precipitate was removed by filtration, washed with 20 ml of dry acetone, air dried and then at 50-55 ° C under vacuum overnight. The glausine phosphate product (yield 1.0 g) melted at 20 DEG-2 DEG. C, H, N found: 51.2, 6.07, 2.96; calculated for ^ 21 ^ 25 ^ ¾ ^^^ ¾ 5 ^ 0.2, 5.91, 2.79 · By the same procedure, using 0.6 gd, 1-glausine in 20 ml of acetone and 0.1 * g of phosphoric acid , the resulting washed, air-dried product was dried under vacuum at 60 ° C for about 2.5 days. Glausine phosphate melted at 2 → 0-2 ^ 2 ° 0. C, H, N found: * + 9.3, 5.91, 2.76; C, H, N calculated for C 21 H 25 N (+, δ ^ ΡΟ ^: b9.2b, 5.86, 2.73- H. 2.5971 kg (5.95 moles) of d, l-glausine hydro bromide, 10.0 liters of deionized water and 3.5 liters of methylene chloride, the mixture was stirred rapidly and 500 ml of 50% sodium hydroxide was slowly added to the solution, and sodium hydroxide was rinsed to 16,681 3,100 ml of deionized water. the mixture was stirred for 15 minutes, then the stirring was stopped and the mixture was allowed to stand for 10 minutes to separate the layers.The methylene chloride layer was discarded and stored.The aqueous layer was stirred with 3.5 liters of methylene chloride and the mixture was stirred rapidly for 15 minutes. allowed to stand for 10 minutes to allow the layers to separate.The methylene chloride layer was discarded.Another 200 mL of methylene chloride was added to the aqueous layer.The methylene chloride layer was discarded. the layers were combined and mixed with 3 liters of deionized water.

The resulting mixture was stirred rapidly for 15 minutes, then allowed to stand for 15 minutes to separate the layers. The methylene chloride layer was discarded and stored. This solution of d, 1-glausine base in methylene chloride was then added to a well stirred solution of 1.4235 kg (12.35 moles) of 85% phosphoric acid in 9.8 liters of toluene-denatured absolute ethanol. A copious white precipitate formed. The precipitate was stirred for 15 minutes, after which it was allowed to stand under a nitrogen atmosphere for about 14-16 hours. Stirring was then started and the precipitate was slowly poured into 3-liter sintered glass funnels. The remaining solid was placed in large, glass drying vessels and air dried, then dried in vacuo at 50-65 ° C to give 2.902 kg (97.1% yield) of d, 1-gl a usinif osf aa tt ia.

A 22 liter flask was charged with 1,500 kg of d, 1-Glucine phosphate and 15 liters of 80% toluene-denatured, absolute ethanol (20% water). The mixture was stirred and refluxed (78 ° C) under nitrogen. The precipitate was refluxed for 5-6 hours, then allowed to cool to 22-25 ° C. The precipitate was then slowly poured into 3 liter sintered soups. The resulting solid was dried. The solid was washed thoroughly with 3 liters of toluene-denatured absolute ethanol and air dried again. The solid was then dried in vacuo at 50-65 ° C to give 1.375 kg (91.7% yield) of d, l-glausine phosphate, which is in a particularly desirable crystalline form in the form of thick slices, i.e. "chunky" as prismatic crystals.

In a differential separation calorimeter, a single peak was obtained for the product, m.p. was 253 ° C. C, H, N found 50.2, 5.97, 176881 3 2.67; C, H, N calculated for c 21 H 25 NO 4 · 1.5H 3 PO 4: 50.2, 5.91, 2.79.

I. 1-Gllausine phosphate prepared as described above was recrystallized three times from ethanol to give the purified salt as fine powdery crystals. C, H, N found: 50.20, 5.91, 2.73; C, H, N calculated for 1.5H 3 PO 4: 50.2, 5.91, 2.79.

The crystal density of J. d, 1-glausine phosphate was determined by suspending at least four crystals of dJ-glausine-1H-H 2 PO 3 in a solvent mixture of benzene and carbon tetrachloride; adjusting the ratio of benzene and carbon tetrachloride to the same density as the suspended crystals; and the density of the solvent mixture obtained from the suspension of the same density was measured using a pycnometer. The crystal density thus obtained was 1.460 g / cm.

The unit cell constants of the crystals of d, 1-glausine'1, δΗ ^ ΡΟ ^ were measured by single-crystal X-ray crystallography. The dimensions of the elemental box were a = 89,854 Angstrom units; b = 8.565 Angström units and c = 23.830 Angström units, the crystals were monoclinic with a β-angle of 93.7 °. Theoretical densities were calculated using the smallest apparent intention volume V = 1/8 abc sin / 3 = 2.286 cubic Angström units (2.286 x 10 "^ cm ^) and Z equal to four molecules / small target field size. For glausine phosphate (1: 1) calculated theoretical density

molecular weight = g Z

- x -

Avogardo Chapter V

O

was 1.319 g / cm. Glausine diphosphate salt 1 e (1: 2) theoretical

O

the density was 1.602 g / cm. The theoretical density of the diglausine monophosphate salt 1a (2: 1) is greater than 2.3 g / cm. For d, 1-glaucine · 1.5H 2 PO 2, the theoretical calculated crystal density was 1.460 g / cm 3, which corresponds to the observed crystal density.

Claims (8)

18 6881 3 A process for the preparation of a therapeutically effective phosphate salt which is 1-glausine phosphate, d, 1-glausine phosphate or a mixture thereof, characterized in that a compound which is 1-glausine, d, 1-g 1 auxin or a mixture thereof is obtained in the form of a base react with excess phosphoric acid. The method of claim 1, wherein the free base, glausine, is dissolved in an inert, organic solvent at a temperature between ambient temperature and the boiling point of the mixture, and the resulting solution is mixed with excess phosphoric acid. The method of claim 1, wherein about 0.5 to 1 mole or two or three molar excess or more of phosphoric acid is used. The process for preparing a phosphate salt according to claim 1, which is 1-gllausine phosphate, d, 1-g 1 auine phosphate or a mixture thereof having about 1.5 molar parts of phosphoric acid per molar part of Glaucine. The process for preparing a d, 1-glausine phosphate salt according to claim 1, having about 1.5 molar parts of phosphoric acid per molar part of d, 1-glausine. The process for preparing a 1-glaucine phosphate salt according to claim 1, which has about 1.5 molar parts of phosphoric acid per molar part of 1-glaucine. Process for the preparation of a phosphate salt which is 1-glausine phosphate, d, 1-glausine phosphate or a mixture thereof according to claim 1, characterized in that a compound which is 1-glausine, d, 1-glausine or a mixture thereof is reacted in the form of a free base with at least a 1.4 molar excess of phosphoric acid,. said phosphate salt has a melting point of about 240-245 ° C with decomposition and a pH of about 2.4-2.6 in aqueous solution (0.5 g / 100 ml). Process for the preparation of a phosphate salt according to claim 1, characterized in that the process has been applied to obtain d, l-glausine phosphate in the form of thick chunky prismatic crystals.