FI69069B - PROCEDURE FOR THERAPEUTIC USE OF THERAPEUTIC NUTRITION N-YAN-N '- (2 - ((4-METHYL-5-IMIDAZOLYL) METHYLTHIO) ETHYL) -N "ALKYNYL LGANIDINER - Google Patents

Description

\ 9λ ** ^ 7 \ γβί Μ1. NOTICE OF PUBLICATION 6906 9 LBJ ιΊΊ) utlAggningsskrift (45) Pa t: L:. ::.: Ε] *: ^ ^ '(51) Kv.lk.4 / lncQ.4 C 07 D 233/64 SUQM | FINLAND (21) Patent application - Patentansökning 781737 (22) Application date - Ansökningsdag 31.05.78 (^) (23) Starting date - Giltighetsdag 31.05.78 (41) Published public - Blivit offentlig 04.12.78

National Board of Patents and Registration Date of publication and publication. - 30.08.85

Patent and registration authorities Ansökan udagd och utl.skriften publicerad (32) (33) (31) Privilege claimed - Begärd priority 03.06.77 22.08.77, 07.11.77 USA (US) 803009, 826796, 848959 (71) Bristol- Myers Company, 345 Park Avenue, New York, New York 10022, USA (72) Ronnie Ray Crenshaw, Dewitt, New York,

George Michael Luke, LaFayette, New York, USA (74) Oy Koister Ab (54) Method for the preparation of therapeutically useful N-cyano-N '- {2 - / (4-methyl-5-imides) ) for the preparation of methyl io_ / ethyl 1 ij -N1'-a 1 kynyyi i Guan id i in ie - ^ Förfarande för framstä11 Ning av therapeutiskt användbara N - cyan-N 1 - ^ - / ^ - mety 1 -5 “ This application relates to certain N-cyano-N '- {2 - (4-methyl-5-imidazolyl) methylthio / ethyl' (-N "-alkynylguanidines and imidazol). , which are I 1 receptor blocking agents that inhibit gastric acid secretion and are useful in the treatment of ulcers.

In the treatment of patients with autoimmune ulcers, the clinical goal is to reduce the secretion of gastric acid, based on the principle of "no acid, no ulcer". The traditional way to treat self-digestive ulcer disease involves dietary control and the use of acid-binding and anticholinergic agents.

There is evidence that histamine may be the ultimate topic for accelerating gastric acid secretion. This effect of histamine is mediated by H2 receptors and is not blocked by classical antihistamines, which are receptor blockers. Several specific β-receptor blocking agents (H2-receptor antagonists) are currently known in 69069. These compounds inhibit basal acid secretion, as well as acid secretion caused by other known gastric acid secretagogues, and are useful in the treatment of self-induction ulcers.

Histamine 1β receptor antagonists are described, for example, in the following patents: U.S. Patent No. 3,950,333 discloses inhibitors of the formula xi— / ΓΛ s v U C- (CH-), Y (CH_) NHCNHR.

X_ ✓ 2 k 2 m l

2 A -— X

wherein A together with the carbon atom is an unsaturated heterocyclic ring having at least one N atom and optionally additionally an S and / or O atom, e.g. (lower) alkyl, X 2 is e.g. hydrogen, k = 0-2, m = 2 or 3 and k + m = 3 or 4, Y is O, S or NH, E is NR 2, wherein R 2 is hydrogen nitro or cyano, and R 1 is hydrogen, (lower ) alkyl or di (lower) alkylamino (lower) alkyl.

BE Patent 804,144 describes compounds of the formula

NCN

HET- (CH-) Z (CH_) NHCNHR.

2 m 2 n 1 wherein HET is a 5- or 6-membered heterocyclic ring containing nitrogen and which may be substituted e.g. with alkyl, m and n are 0-4, m + n = 2-4, Z is S, O, NH or CH 2, and R 1 is H or (lower) alkyl.

BE Patent 841,814 discloses compounds of the formula

X

HET-CH 2 Z (CH 2) 2NHCNHY

where HET is e.g. imidazole, which may contain e.g. alkyl substituent, Z is S or CH 2, x is S, CHNC> 2, NCN or NH, Y is NH 2, (lower) alkylamino, di (lower) alkylamino, (lower) alkoxy, phenylethyl, imidazolylethyl, allyl , trifluoroethyl or (CH 2> n R, n = 1-12 and R is OH, (lower) alkoxy, NH 2 or (lower) alkylamino,

II

69069 wherein when X is NH Y is trifluoromethyl or (CHO) R, and when X is NCN Y is not amino or (lower) alkyl.

U.S. Patent No. 3,894,151 describes a variety of groups of compounds for use as H 2 -histamine inhibitors.

BE Patent 843,840 discloses compounds of the formula

X

CH_Z (CH2) NHC-NHY N- / 2 2 n t 1

H CH2OH

where n = 2-3, Z is S or CH 2, Y is e.g. (lower) alkyl and X is e.g. NCN.

The present invention relates to a process for the preparation of therapeutically useful N-cyano-N '(4-methyl-5-imidazolyl) methyl-η-thio_7-ethyl] -N "-alkynylguanidines of the formula jr h N 3 NCN 1

\ "1 \ N / CH2SCH2CH2NHCNH-R H

wherein R 1 is a straight or branched C 1 -C 4 alkynyl group, preferably 4 4 J 4 -CHC = CR or "<CH 2 ^ n = CR '30; * in which R is H or CH 2 and ch 3 n is 1-6 , and pharmaceutically acceptable acid addition salts thereof.

The process according to the invention is characterized in that a) a compound of the formula CH, SI- ^ 3 | vii '' / 'ay

B

or an acid addition salt thereof, wherein X is a leaving group, preferably halogen, is reacted with a compound of formula 4 69069

NCN

II 1

HSCH2CH2NHCNH-R VIII

wherein R is as defined above, or b) a compound of formula N-

I II

CHR SCHRCH NHRr H 1 or an acid addition salt thereof is reacted with a compound of formula

R, NHR1 III

o in which formulas R has the same meaning as above and one of the symbols Rj and Rg is hydrogen and the other a group of formula

NCN

il

-C-SR

wherein R is a substituent such that -SR is a leaving group, and the resulting compound of formula I is optionally converted to a pharmaceutically acceptable acid addition salt.

Preferred compounds of the invention are N-cyano-N '- (2 - [' (4-methyl-5-imidazolyl) methylthio] ethyl) -N "- (2-butyn-1-yl) guanidine, N-cyano-N ' - (2- [(4-methyl-5-imidazolyl) methylthio / ethyl) -N "- (3-butyn-1-yl) guanidine, N-cyano-N '- (2-Z (4-methyl- 5-imidazolyl) methylthiomethyl) -N "- (4-pentynyl-1-yl) guanidine, N-cyano-N '- (2- (4' - (4-methyl-5-imidazolyl) methylthio] ethyl) -N" - (3 -butyn-2-yl) guanidine and their non-toxic pharmaceutically acceptable acid addition salts.

69069

A particularly preferred compound is N-cyano-N '- (2-Z "(4-methyl-5-imidazolyl) methylthio / ethyl) -N" -propargylduanidine and its non-toxic pharmaceutically acceptable acid addition salts.

The preparation of this preferred compound is illustrated in Reaction Schemes I and II.

Scheme I CH

N ^

NCN

^ CH2SCH2CH2NHC-SR + H2NCH2C2CH H

V

_ / CH3 N Γ

J 1 NCN

\ N Il N CHoSCH0CHoNHCNHCH „C = CH H 2 2 2 2

The reaction is carried out in an inert solvent at a temperature higher than room temperature. As will be apparent to one skilled in the art, R may be any substituent such that -SR is a suitable leaving group. Thus, R may be (lower) alkyl, aryl, substituted aryl (e.g., p-nitrophenyl), aralkyl, -CH 2 CN, -CH 2 COOH, -CH 2 COOR ', or the like. N-Cyano-N '- (2 - [' (4-methyl-5-imidazolyl) methylthio / ethyl) -SR-isothiourea starting materials can be prepared by the methods described in BE Patent 804 144. The alkynylamine starting materials are either commercially available. substances or can be prepared by the methods described in Bull. Soc. Chim. Fr. 490 (1958), Bull. Soc. Chim. Fr., 592 (1967) and Annales de Chimie (Paris), 3, 656 (1958).

6

Figure II

69069 M ^ CH, N --- f— 3

U i NCN

\ il xch2sch2ch2nh2 + rs-cnhch2c = ch

B

\ / N - ^ CH3

j I

NCN

\ ^ N II

N CH2SCH2CH2NHCNHCH2C2CH

The reaction is carried out in an unreacted solvent at a temperature higher than room temperature. The substituent R may be the substituent mentioned in Scheme I above. The 2- [4- (4-methyl-5-imidazolyl) methylthio] ethylamine starting material can be prepared as described in U.S. Patent 3,950,353. The di-substituted cyanodithioiminocarbonate used as a starting material in the preparation of N-cyano-N'-propargyl-SR-isothiourea (see step b of Example 2) can be prepared by the methods described in J.Org. Chem., 32, 1566 (1967).

It has further been found that in the preparation of N-cyano-N '- (2 - [' (4-methyl-5-imidazolyl) methylthio] ethyl) -N "-propargylguanidine according to reaction formula I, preferably by selecting the reaction conditions of the last step of the process, the product Under the latter conditions, the crude product can be isolated as a crystalline solid which can be purified more thoroughly by simple recrystallization, thus avoiding chromatographic purification of the crude product, which was previously considered to be the most preferred other. when using reaction conditions.

Methanol is used as a solvent in the above reactions. The reaction is carried out in a concentrated solution (about 1 g of substituted isothiourea per 5 ml of methanol) under a nitrogen atmosphere and using freshly distilled propargylamine.

Experiments have shown that the use of a nitrogen purge prevents the formation of two hitherto unidentified by-products, which by-products are formed when nitrogen is not used. These by-products could not be removed by column chromatography or recrystallization. It is believed that nitrogen purging prevents the formation of these by-products by removing mercaptan gas at the same rate as it is formed.

The reaction of process variant a) is carried out in an inert organic solvent. In a preferred embodiment, the compounds of formulas VII and VIII are used in equimolar amounts, and the reaction is carried out in the presence of a base.

The leaving groups "X" suitable for use in this reaction are well known to those skilled in the art. These are, for example, fluorine, 2 2 chlorine, bromine, iodine, -O-SR, where R is (lower) alkyl / e.g. methanesulfonate /, O 3 SR, where R is aryl or substituted aryl / e.g. benzenesulfonate, p-bromobenzenesulfonate or p-toluenesulfonate_7, O, SF, acetoxy and 2,4-dinitrophenoxy. For convenience and economic reasons, a compound of formula VII wherein X is chlorine is most preferably used.

In this method, the compound of formula VII is used mainly in the form of an acid addition salt. Compound VII is relatively unstable as the free base and is therefore prepared and stored in the form of an acid addition salt. Although the compound of formula VII can be converted to a base before carrying out the reaction, no advantage is obtained in this process. It will be apparent to those skilled in the art that any inorganic acid or organic acid can be used to form the acid addition salt of a compound of formula VII, e.g. the compound of formula VII is usually used in the form of the hydrochloride salt.

8 69069

The reaction between compounds VII and VIII to prepare a compound of formula I can be carried out in any inert organic solvent such as alkanol, acetonitrile, dimethylformamide, dimethyl sulfoxide, acetone and the like. Preferred solvents are alkanols such as ethanol or 2-propanol.

The reaction temperature is not critical; the reaction can be carried out at temperatures between about 0 and about 200 °. At low temperatures the reaction is slow, while at high temperatures less pure products are formed due to decomposition and formation of by-products. In general, we find it convenient to carry out the reaction at room temperature.

The reaction between compounds VII and VIII to prepare compound I is carried out mainly in the presence of a base, which facilitates the reaction when the base acts as an acid scavenger. Suitable bases for use in this reaction include both inorganic and organic bases such as NaOH, KOH, LiOH, triethylamine, dimethylaniline, sodium ethylate and the like.

The intermediate of formula VIII NCN H 1 used in the process of the invention

hsch2ch2nhcnh-r 'VIII

wherein R is a straight or side chain alkynyl group having 3 to 9 carbon atoms is a novel compound.

1 4 4

In the formula Vili R is preferably -CHC = CR, where R is hydrogen

CH

4 4

or methyl or - (CH 2) C-CR, where n is an integer from 1 to 6 and R is

2 n is hydrogen or methyl ;.

Examples of preferred meanings of R 1 in formula VIII are 2-butyn-1-yl, 3-butyn-1-yl, 4-pentynyl-1-yl. The most preferred meaning is a propargyl group.

Intermediates of formula VIII li 69069 NCN 11 1

HSCH2CH2NHCNH-R VIII

is prepared by reacting a compound of formula

p-sch2chnh2 IX

wherein P is any group suitable as a sulfhydryl protecting group to react with a compound of formula

NCN

II

RSCNCH2C = CH X

and removing the sulfhydryl protecting group of the resulting compound.

The dimethyl cyanodithioiminocarbonate used as a starting material in the preparation of N-cyano-N'-alkynyl-S-methylisothiourea compounds can be prepared by the method described in J. Org. Chem., 32, 1566 (1967). Alkynylamine starting materials are either commercially available or can be prepared by the methods described in Bull. Soc. Chim. Fr., 490 (1958); Bull. Soc. Chim. Fr., 592 (1967) and Annales de Chimie (Paris), 2, 656 (1958).

Histamine H2 receptor antagonists have been shown to be effective inhibitors of gastric acid secretion in animals and humans, Brismblecombe et al., J. Int. Med. Res., 3, 86 (1975). A clinical study of the histamine H2 receptor antagonist cimetidine has shown it to be an effective therapeutic agent in the treatment of autoimmune ulcer disease, Gray et al., Lancet, 1 (8001), 4 (1977). The compound prepared in Examples 1 and 2 below (hereinafter referred to as BL-5641) has been compared with cimetidine in various experiments and has been found to be much more effective than cimetidine both as a histamine H2 receptor antagonist in isolated guinea pig atria and as an inhibitor of gastric acid secretion in rats and dogs. In addition, gastric acid secretion experiments in dogs show that the duration of action of BL-5641 at longer doses is longer than that of cimetidine.

1 o 69069

Histamine H2 receptor antagonism -

Experiment performed in an isolated guinea pig atrium

Histamine causes a concentration-dependent increase in the rate of contraction in the isolated, spontaneously striking right atrium of a guinea pig. Black et al., Nature, 236, 385 (1972), call receptors with this form of histamine action histamine H2 receptors when reporting the properties of burimamide, which always competes as an antagonist of these receptors. Subsequent studies by Hughes and Coret. Proc. Soc. Exp.

Biol. Med., 148, 127 (1975) and Verma and McNeill, J. Pharmacol. Exp. Ther., 200, 352 (1977), support the conclusion of Black and colleagues that the positive pulse-regulating effect of histamine in the isolated right atrium of a guinea pig is mediated by histamine H2 receptors. Black et al., Agents and Actions, 3, 133 (1973) and Brimble-comge et al., Fed. Proc., 35, 1931 (1976) have used isolated guinea pig right atria as a means of comparing the effects of histamine H2 receptor antagonists. These comparative studies were performed using a variation of the method described by Reinhardt et al., Agents and Actions, 4, 217 (1974).

Male Hartley guinea pigs (350-450 g) were killed by striking the head. The heart was excised and placed in a Petri dish containing oxidized (95% O 2, 5% CO 2), modified Krebs solution (g / liter: NaCl 6.6, KCl 0.35, MgSO 4 · 7 H 2 O 0.295, KH 2 PO 4 0.162, CaCl 2 0.238, NaHCO 2 2.1 and dextrose 2.09).

Other tissues were dissected from the spontaneously pulsating right atrium and a silk thread was tied at each end (4-0). The atrium was suspended in a 20 ml muscle compartment containing an oxidized, modified Krebs solution maintained at 32 ° C. Atrial contractions were recorded isometrically with a Grass FT 0.03 transmission transducer and contraction force and velocity recordings were performed on a Beckman RP Dynograph.

A resting tension in the atrium was applied to a force of 1 g and allowed to equilibrate for one hour. At the end of the equilibration period, almost the maximum amount of histamine dihydrochloride (3 x 10 6) was added to the bath and washed to embed the tissue. Histamine was then added to the bath in increasing amounts using 1/2 log 10 intervals to give final bath molar concentrations of --7 -5 to 1x10 -3x10. The histamine-induced atrial velocity of 11,69069 was allowed to stabilize before the next successive increase.

The maximum reaction occurred unchanged at a concentration of -5 3x10 M. The histamine was washed off several times and the atrium was allowed to return to the control rate. Test compound -5 (1 x 10 M) was then added and after a germination time of 30 minutes, the histamine concentration reaction was repeated by adding higher concentrations as needed.

Histamine ED50 values (concentration of histamine that increased the rate of contraction by 50% of the maximum) and 95% confidence limits before and after addition of test compound were obtained by regression analysis as described by Finney, Probit Analysis, 3rd ed., Cambridge (1971). The transition coefficients of the concentration-response curve were calculated as follows: ED50 histamine + compound

Transition coefficient = - ED50 histamine The coefficient obtained with BL-5641 alone was then expressed as the ratio of the coefficient obtained with cimetidine.

BL-5641 birth rate -1

Activity Ratio = - Cimetidine Shift Coefficient -1 The results obtained in these studies are summarized in Table 1. Cimetidine and BL-5641 shifted the histamine-concentration-response curve to the right with coefficients of 6.6 and 32.7, respectively. Based on the transition coefficients of the concentration-response curve, BL-5641 was approximately 5.7-fold more active than cimetidine as a histamine H2 receptor antagonist in the isolated guinea pig right atrium.

69069 <1 2

table 1

Cimetidine and the relative activity of BL-5641 in an isolated guinea pig right atrium

Histamine ED50 Concentration Active

Compound N Concentration in reactant ratio (95% Oug / ml). transition period _____roin__

Histamine 0.21 control 3 (0.18 - 0.25) - --- 1.39

Cimetidine 3 1 x 10-5M (1.08-1.85) 6.6 1.0

Histamine 0.38 control 2 (0.27-0.53) 12.44 BL-5641 2 1 x 10 "5M (7.81-20.28) 32.7 5.7 N = number of experiments.

Determination of the inhibitory effect of gastric acid secretion in rats with the gastric port bound for two hours (Shay) The gastric port binding procedure in rats was designed by Shay et al., Gastroenterology, 5, 53 (1945) for gastric ulcer outbreak studies; whereas, however, once the method became known, it was used as a means of studying gastric acid secretion in rats;

Shay et al., Gastroenterology, 26, 906 (1954), Brodie, D. A., Am. J.

Dig. Dis., 11, 231 (1966). A variation of this method has been used without delay to evaluate the ability of the compounds to inhibit the secretion of the antacid.

Long Evans male rats, 280-300 g, have been used in the experiments. The animals are placed in separate cages and fasted for 24 hours with free water. Under ether anesthesia, the abdomen is accessed by opening down to the midline and a cotton bandage is placed around the gastric port. After closing the wound with ether

II

13 69069 is discontinued and either BL-5641, cimetidine or medium is administered intraperitoneally at 1 mg / kg. BL-5641 and cimetidine are dissolved in one equivalent of HCl and made up to volume with water. The animals are returned to their cages from which the water bottles have been removed and two hours later they are killed with ether. The stomach is removed and the two-hour gastric suture is drained into a graduated test tube to determine volume. The titratable acidity is measured by titrating a 1 ml sample to pH 7.0 with 0.02 N NaOH using an automatic burette and a potentiometric pH meter (radiometer). The liberated acid to be titrated is calculated in microequivalents by multiplying the volume in milliliters by the acid concentration in milliequivalents per liter. The percentage inhibition of acid release is calculated as follows:

Acid release inhibition% = released acid control - released acid - drug χ ^ qq released acid control The results obtained with BL-5641 and cimetidine are shown in Table 2.

These results indicate that in a preparation using a two-hour bound rat gastric port, BL-5641 was at least as effective in inhibiting gastric acid formation as cimetidine.

14 69069

Table 2 Effect of BL-5641 and cimetidine on gastric acid formation in the rat gastric port bound for 2 hours

Compound Dose (ip) a ED50 of acid formation _MJ moles / kg mg / kg inhibition% __ / jmolesAg BL-5641 40 11.1 80 20 5.53 61 ~ 11 10 2.76 37

Cimetidine 40 10 66 20 5 60 rs »14 10 2.5 40 5 1.25 34 a At least five animals were used for each dose.

Determination of the inhibitory effect of gastric acid secretion in the abdominal cavity of a dog

Thomas-type Thomas, J.E., Proc. Soc. exp. Biol.

Med., 46 260 (1944) _7 stainless steel cannulas are placed in the stomachs of hare dogs (10-12 kg) just at the glandular region of the gastric port near the major gastric arch to provide a long-term abdominal opening. The animals are allowed to recover for at least two months before any experiments are performed. However, before each experiment, dogs are fasted overnight (»~ 18 hours) with free water. Dogs are tethered and an eight-inch internal needle catheter (C. R. Baird, Inc.) with a two-inch 17-gauge needle is inserted into the leg vein for drug administration. Gastric secretions are collected in a coca for 15 minutes by draining from an opened cannula. Basal secretions are pooled from two consecutive 15-min batches and if these prove to be too large (> 4 ml / 15 min; pH <5.0), animals are not used. The method variation described by Grossman and Konturek, Gastroenterology, 66, 517 (1974) was followed. Immediately after collection of the second batch, histamine (100 ug / kg / h) is injected for 90 minutes with a 6 ml / h Harvard Infusion Pump. At this time, li 15 69069 is injected rapidly (within 30 seconds), 0.1 mg / kg of either BL-5641, cimetidine (solubilized with one equivalent of HCl and made up to volume with normal saline), or ordinary saline, followed by injection of histamine. continues for another 150 minutes (total spray time is 4 hours). Each 15-minute gastric fluid sample is measured more closely at 0.5 ml and the titratable acidity against 0.02 N NaOH (end point pH 7.0) is measured using an automatic burette and a pH meter (radiometer). The percentage inhibition of acid secretion is calculated as described in connection with a method using a rat-bound gastric port.

Five different dogs were given equimolar doses of BL-5641 and cimetidine and the results obtained are shown in Table 3.

Both BL-5641 and cimetidine had an immediate inhibitory effect on gastric acid secretion. At equimolar doses, BL-5641 consistently had a higher percent inhibition and a longer duration of action than cimetidine. These results indicate that BL-5641 is more potent and / or longer acting than cimetidine as an inhibitor of histamine-induced gastric acid formation in the body.

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The following examples illustrate the invention.

Example 1 N-Cyano-N '- (2- (4') - (4-methyl-5-imidazolyl) methylthio] -ethyl-N "-propargylguanidine / CH 3 N"

11 NCN

\. '\ Il

^ N 'λCH2SCH2CH? NHC-SCH3 + H2NCH2C = CH

V

/ CH3 N-

l * i NCN

\ N I »

H CH2SCH2CH2NHCNHCH2C = CH

A mixture of N-cyano-N '- (2-Z- (4-methyl-5-imidazolyl) methylthio] ethyl) -S-methylisothiourea (3.00 g, 0.0111 mol) and propargylamine ( 2.50 g, 0.045 mol) in acetonitrile (60 ml) was stirred at reflux for 65 hours and then heated in a stainless steel pressure vessel at 120-130 ° for 38 hours. The reaction mixture was cooled, decanted apart from the tar, and then evaporated to dryness to leave a resin (3.57 g). This material was taken up in silica gel (100-200 mesh) and eluted with a mixture of methylene chloride (97 parts) and methanol (3 parts). The product from the intermediate fraction was crystallized by trituration under acetonitrile, and then recrystallized from acetonitrile to give the title compound (0.236 g; 7.7%); mp. 146 to 149.5 °.

and 18,69069

Analysis calculated for C 12 H 16 N 6 S: C, 52.15; H, 5.83; N, 30.41; Found: C, 51.86; H, 5.81; N, 30.70.

Example 2 N-Cyano-N '- (2- [~ (4-methyl-5-imidazolyl) methylthio] ethyl) -N "-propargylguanidine (ch3s) 2c = ncn + h2nch2c = ch n /

NCN

II

CH3SCNHCH9C = CH

A

CH0 N - / h i n "\

nCH2SCH2CH2NH2 + A

V

CK

OF-

Jl NCN

^ N '\ Il

„CH ^ SCHnCH-NHCNHCHOC = CH M L · Z Z Z

19 69069 a. N-Cyano-N'-propargyl-S-methylisothiourea (A)

A solution of dimethyl cyanodithioiminocarbonate (16.00 g, 0.109 mol) and propargylamine (6.03 g, 0.109 mol) in acetonitrile (320 ml) was stirred at reflux for 4 hours and then at 25 ° for 12 hours. . Further work-up gave the title compound A (13.58 g 81%), m.p. 160-164 °.

b. N-cyano-N1- (2- [(4-methyl-5-imidazolyl) methylthio] -ethyl-N "-propargylguanidine

A solution of Compound A (11.71 g, 0.0765 mol) and 2 - [(4-methyl-5-imidazolyl) methylthio] ethylamine (13.10 g, 0.0765 mol) in methanol (250 ml) was heated to reflux. 64 hours. The solvent was removed by evaporation to dryness, the residue was added to silica gel (100-200 mesh) and chromatographed using gradient elution with methylene chloride / methanol; 4.0 g of the title compound were obtained from the latter fractions. Recrystallization from acetonitrile gave the pure product, m.p. 150-152.5 °, which was identical (ir, nmr, tie) to the product prepared in Example 1.

Example 3 N-Cyano-N '- (2 - [' (4-methyl-5-imidazolyl) methylthio] ethyl) -N "- (2-butyn-1-yl) guanidine (CH, S)" C = NCN + HnNCH »C-CCH ^ 3 2 δ 2 3

NCN

H Ύ CH 3 SCNHCH 2 C = CCH 3

B

_____ ί! 20 69069 CH ^ N - ^ I i I i ^ \

H CH2SCH2CH2NH2 + B

! j Ψ CH3 N- i * Ji \ νΛ!? cn H CH-SCH „CH ^ NHCNHCH-C2CCH-, 2. 2. δ 2 3 a. N- (2-butyn-1-yl) -N1-cyano- S-methyl isothiourea (B)

A solution of dimethyl cyanodithioiminocarbonate (10.00 g, 0.0684 mol) and 2-butyn-1-ylamine (4.73 g, 0.0684 mol) in acetonitrile (200 ml) was stirred at 25 ° for 0.5 hours, and then boiled for 2.5 hours. The mixture was cooled, then filtered to give the title compound B, m.p. 180-183 °.

b. N-cyano-N1- (2- [4- (4-methyl-5-imidazolyl) methylthio] ethyl) -N "- (2-butyn-1-yl) guanidine

A solution of compound B (6.82 g, 0.0407 mol) and 2-Z ”(4-methyl-5-imidazolyl) methylthio] ethylamine (6.98 g, 0.0407 mol) in methanol (140 ml) was heated to reflux. 40 hours. After further work-up and chromatography as described in Example 2 above, the title compound was obtained. When the oyster compound was prepared according to the general method of Example 1, it melted at 128-130 °.

Example 4 N-Cyano-N '- (2- [(4-methyl-5-imidazolyl) methylthio] ethyl) -N "- (4-pentynyl-1-yl) guanidine

The general procedure of Example 1 is repeated, but the propargylamine used therein is replaced with an equimolar amount of 4-pentyn-21,69069 1-ylamine to give the title product; mp. 99-103 °.

Example 5 N-Cyano-N '- (2 - [(4-methyl-5-imidazolyl) methylthio] ethyl) -N "- (2-butyn-1-yl) guanidine

A mixture of N-cyano-N '- (2 - [' (4-methyl-5-imidazolyl) -methylthio] -ethyl) -5-methylisothiourea (3.00 g, 0.0111 mol) and 2- butyn-1-ylamine (3.07 g, 0.0445 mol) in 60 ml of propionitrile was stirred at reflux for 40 hours. Plate chromatographic analysis of an aliquot sample of the reaction mixture showed traces of the isothiourea starting material, so the mixture was boiled for another 6 hours and then stirred for 64 hours at room temperature. The solvent (and with it the excess amine) was removed in vacuo and the remaining resin was added to silica gel (100-200 mesh) and eluted with a mixture of methylene chloride (97 parts) and methanol (3 parts). The intermediate fractions were combined and evaporated to dryness to give 1.81 g of a yellow resin. The resin was dissolved in 20 ml of ethyl acetate and crystallized at -15 ° C. The resulting pale yellow solid (1.2 g) was dissolved in 11 ml of hot acetonitrile and recrystallized at -15 ° C; yield 1.072 g, m.p. 128-130 ° C.

Analysis calculated for C 18 H 18 N 2 S: C, 53.77; H, 6.25; N, 28.94; S, 11.08 Found: C, 53.72; H, 6.29; N, 29.62; S, 11.34

Example 6 N-Cyano-N '- (2 - [' (4-methyl-5-imidazolyl) methylthio] ethyl) -N "- (3-butyn-1-yl) guanidine

A mixture of N-cyano-N '- (2 - [' (4-methyl-5-imidazolyl) methylthio / ethyl) -5-methylisothiourea (3.00 g, 0.0111 mol) and 3-Butyn-1-ylamine (3.13 g, 0.0453 mol) in 60 ml of propionitrile was stirred at reflux for 40 hours. The solvent was evaporated to give 5.40 syrup which was added to 70 g of silica gel (100-200 mesh) and chromatographed using gradient elution with methylene chloride / methanol.

22 6 9 0 6 9

Fraction Elution mixture Volume Weight 1 99 CH 2 Cl 2/1 MeOH 100 ml 2 "70 ml 3" 50 ml 0.117 g 4 "30 ml 5" 75 ml 0.067 g 6 "150 ml 0.080 g 7" 325 ml 0.168 g 8 98 CH2Cl2 / 2 MeOH 50 ml 9 "150 ml 0.125 g 10" 250 ml 0.811 g 11 "250 ml 0.72 g 12 97 CH2Cl2 / 3 MeOH 175 ml 0.125 g 13" 225 ml 0.133 g 14 "- 0.027 g 15" 225 ml 1 f 0.002 g

16 "200 mlJ

17 95 CH 2 Cl 2/5 MeOH 250 ml 0.035 g 18 "250 ml 0.236 g

Plate chromatographic analysis (silica gel, 90 CH 2 Cl 2/10 MeOH) showed that fractions 9-12 were pure and could be combined. Fractions 1-8 showed rapidly moving impurities. Fractions 13-14 showed some entrained impurities. Fractions 9-12 were combined and evaporated to 1.72 g of a yellow resin. The resin was dissolved in 11 ml of nitromethane and crystallized at -15 ° C; a pale yellow solid, 1.18 g, which was recrystallized from 9 ml of nitromethane to give 0.987 g of product; mp. 86-89 ° C (soft at 85 ° C. Infrared and NMR (100 MHz) were pure and consistent with the desired structure. NMR indicated that the product was solvated with approximately 0.08 moles of nitromethane.

Analysis calculated for C 18 H 18 N 2 S. 0.08 (CH 2 Cl 2): C, 53.21; H, 6.22; N, 28.84; S, 10.86; Found: C, 52.68; H, 6.28; N, 29.39; S, 11.22.

52.87; H 6.02; N, 29.50.

I! 23 69069

Example 7 N-Cyano-N * - (2 - [(4-methyl-5-imidazolyl) methylthio] -ethyl) -N '- (4-pentynyl-1-yl) guanidine

A mixture of N-cyano-N '- (2- [4' - (4-methyl-5-imidazolyl) methylthio] ethyl) -5-methylisothiourea (3.00 g, 0.0111 mol) and 4- pentyn-1-ylamine (3.69 g, 0.0455 mol) in 60 ml of acetonitrile was stirred at reflux for 24 hours and then allowed to stand at room temperature for 96 hours. The solvent and excess amine were removed in vacuo and the remaining yellow resin was purified by chromatography on 50 grams of 100-200 mesh silica gel using a methylene chloride / methanol mixture (99: 1 to 96: 4) in a gradient elution. The intermediate fractions, which were found to be pure by plate chromatography, were combined and evaporated to dryness to give 1.91 g of a yellow resin which was crystallized at -15 ° C from 18 ml of ethyl acetate. The resulting white solid (1.25 g) was recrystallized at -15 ° C from 10 ml of acetonitrile to give 1.063 g of product, m.p. 99-103 ° C.

Analysis calculated for C 14 H 20 N 6 S: C, 55.24? H, 6.52; N, 27.61? S, 10.53? Found: C, 55.43; H, 6.58; N, 28.26? S, 10.97.

Example 8 N-Cyano-N '- (2- [7,4-methyl-5-imidazolyl) methylthio] -ethyl-N "-propargylguanidine

A mixture of N-cyano-N '- (2 - [(4-methyl-5-imidazolyl) methylthio] ethyl) -5-methylisothiourea (10.0 g; 0.0371 mol) and distilled propargylamine (20 ml; 0.325 mol) in methanol (50 ml) was stirred and refluxed under positive nitrogen pressure (nitrogen purge) for 20.5 hours, after which the solvent and excess amine were removed by evaporation to dryness to leave a tan oil which crystallized easily. Roasting the crude product under ispropanol (30 mL) gave the title compound as a cream, brittle solid (8.11 g, 79%), mp 146-148.5 °.

24 69069

Example 9 N-Cyano-N '- (2 - [' (4-methyl-5-imidazolyl) methylthio] ethyl) -N "-propargylguanidine

A mixture of N-cyano-N '- (2 - [(4-methyl-5-imidazolyl) methylthio] ethyl) -5-methylisothiourea (100 g; 0.371 mol) and distilled propargylamine (150 ml. 2 , 44 moles) in methanol (500 ml) was stirred and refluxed under positive nitrogen pressure (nitrogen purge) for 22 hours. The reaction mixture was cooled and the solvent and excess amine were removed by evaporation to dryness, leaving a golden brown oil which crystallized easily. The crude product was triturated under isopropanol (250 mL), cooled to 0 ° for 2 hours, and the product was collected by filtration. The filter cake was washed with cold isopropanol and dried under vacuum at Ρ2 <3, η for 16 hours. The dried title product was an off-white, dense, solid; yield 73.5 g (71.7%), m.p. 147-149 °.

Example 10 Recrystallization of N-cyano-N '- (2- [(4-methyl-5-imidazolyl) -methylthio] -ethyl) -N "-propargylguanidine The final products obtained in Examples 11 and 12 were combined (total weight 81.3 g), dissolved to hot isopropanol (1000 mL), filtered through Super-Cel and allowed to cool at room temperature for about 68 hours The resulting crystalline product was collected by filtration, washed with cold isopropanol, finely ground and dried in a heated desiccator under high vacuum for about 45 hours. .4 g (recovered 89%), mp 149-151 ° C. The HPLC assay showed a purity of about 99.5%.

The NMR (100 MHz) spectrum was pure and of uniform quality. Analysis calculated for C 12 H 16 N 6 S: C, 52.15? H, 5.83; N, 30.41; S, 11.60? Found: C, 52.42; H, 5.94; N, 30.51; S, 11.35.

Example 11 N-Cyano-N '- (2 - [' (4-methyl-5-imidazolyl) methylthio7-ethyl) -N "-propargylguanidine A. N-Cyano-N1-propargyl-S-methylisothiourea A solution in which was dimethyl cyanodithioiminocarbonate (16.00 g, 0.109 mol) and propargylamine (6.03 g, 0.109 mol)

II

69069 in acetonitrile (320 ml) was stirred at reflux for 4 hours and then at 25 ° for 12 hours. The mixture was cooled and filtered to give the title compound (13.58 g, 81%) m.p. 160-164 °.

B. N-Cyano-N'-propargyl-N "- (2-mercaptoethyl) guanidine A mixture of 1.136 g (10 mmol) of cysteamine hydrochloride, 1.53 g (10 mmol) of the product of step A above and 0.055 g of hydroquinone in 10 ml of DMF was warmed slightly to give a solution, to which was added 10 ml of 1N aqueous sodium hydroxide solution, and nitrogen was bubbled through the solution. and a mixture of sodium chloride The title product was extracted from this mixture with 10 ml of ethanol and the ethanol solution was used in step C below.

C. N-Cyano-N '- (2- (4') - (4-methyl-5-imidazolyl) methylthio] ethyl) -N "-propargylguanidine

An ethanolic solution of the product of Step B above (ca. 10 mmol) was added to a solution of 0.46 g of sodium (0.02 g atoms) in 10 mL of ethanol under nitrogen at 4 ° C. After 5 minutes, a solution of 1.67 g (10 mmol) of 4-methyl-5-chloromethylimidazole was added. HCl in 14 mL of ethanol and the mixture was stirred at room temperature under nitrogen for 70 minutes. The reaction mixture was filtered through a pad of Celite to remove inorganic salts and the Celite was washed with ethanol. The filtrate was evaporated to dryness and the product was purified by chromatography on a silica gel column (20 g silica, 3.2 x 4.5 cm mattress) using ethanol / chloroform mixtures as solvents. The ethanol content was gradually increased from 2 to 15%. The eluate was evaporated to dryness to give 1.906 g of crystalline product. Recrystallization from 2-propanol gave 1.49 g (54%) of the title compound, m.p. 144-145 ° C.

Example 12 N-Cyano-N '- (2 - [' (4-methyl-5-imidazolyl) methylthio-7-ethyl) -Nn- (2-butyn-1-yl) guanidine A. N-Cyano-N '- ( 2-Butyn-1-yl) -5-methylisothiourea A solution of dimethyl cyanodithioiminocarbonate (10.00 g, 0.0684 mol) and 2-butyn-1-ylaramine (4.73 g, 0.0684 mol) in acetonitrile (200 ml) was stirred at 25 ° for 0.5 hours, then boiled for 2.5 hours. The mixture was cooled and then filtered to give the title compound, m.p. 180-183 °.

B. N-Cyano-N1- (2-butyn-1-yl) -N "- (2-mercaptoethyl) -guanidine

The general procedure of Example 11B is repeated, but the N-cyano-N'-propargyl-S-methylisothiourea used therein is replaced by an equimolar amount of N-cyano-N1- (2-butyn-1-yl) -S-methylisothiourea, and thus the title product is obtained.

C. N-Cyano-N '- (2- [14-methyl-5-imidazolyl) methylthio] ethyl) -N "- (2-butyn-1-yl) guanidine

The general procedure of Example 11C is repeated, but the N-cyano-N'-propargyl-N "- (2-mercaptoethyl) guanidine used therein is replaced by an equimolar amount of N-cyano-N '- (2-butyn-1-yl) -N" - (2-mercaptoethyl) guanidine, to give the title product.

Example 13 N-Cyano-N '- (2- [14-methyl-5-imidazolyl) methylthio] ethyl) -N "- (3-butyn-1-yl) guanidine A. N-Cyano-N' - (3 -butyn-1-yl) -N "(2-mercaptoethyl) guanidine The general procedures of Example 11, steps A and B are repeated, but the propargylamine used in step A is replaced with an equimolar amount of 3-butyn-1-ylamine to give the title product.

B. N-Cyano-N '- (2- [14-methyl-5-imidazolyl) methylthio] ethyl) -N' '- (3-butyn-1-yl) guanidine

The general procedure of Example 11C is repeated, but the N-cyano-N'-propargyl-N "- (2-mercaptoethyl) guanidine used therein is replaced by an equimolar amount of N-cyano-N '- (3-butyn-1-yl) -N" - (2-mercaptoethyl) guanidine to give the title product, m.p. 86-89 ° C.

Example 14 N-Cyano-N '- (2- [14-methyl-5-imidazolyl) -methylthio] -ethyl-N "- (4-pentyn-1-yl) guanidine A. N-Cyano-N' - ( 4-pentyn-1-yl) -N '- (2-mercaptoethyl) guanidine

The general procedures used in Steps A and B of Example 11 are repeated, but the propargylamine used in Step A is replaced with an equimolar amount of 4-pentyn-1-ylamine to give the title product.

li 27 69069 B. N-cyano-N '- (2- [4- (4-methyl-5-imidazolyl) methylthio] ethyl) -N "- (4-pentynyl-1-yl) guanidine

The general procedure of Example 11C is repeated, but the N-cyano-N'-propargyl-N "- (2-mercaptoethyl) -guanidine used therein is replaced by an equimolar amount of N-cyano-N '- (4-pentynyl-1-yl) -N "- (2-mercaptoethyl) guanidine, to give the title product.

Example 15 N-Cyano-N '- (2- [(4-methyl-5-imidazolyl) methylthi-7-ethyl) -N "- (3-butyn-2-yl) guanidine A. N-Cyano-N' - ( 3-butyn-2-yl) -N '- (2-mercaptoethyl) guanidine

The general procedures of Example 11, Steps A and B are repeated, but the propargylamine used in Step A is replaced with an equimolar amount of 1-methylpropargylamine to give the title product.

B. N-Cyano-N '- (2- [4-methyl-5-imidazolyl) methylthio] ethyl) -N "- (3-butyn-2-yl) guanidine

The general procedure of Example 11 is repeated, but the N-cyano-N'-propargyl-N "- (2-mercaptoethyl) guanidine used therein is replaced by an equimolar amount of N-cyano-N '- (3-butyn-2-yl) -N" - ( 2-mercaptoethyl) guanidine to give the title product, m.p. 125-128 ° C.

Claims (2)

A process for the preparation of therapeutically useful N-cyano-N'-4-methyl-5-imidazolylmethylthio] ethyl} -N "-alkynylguanidines of the formula CH3, S - f II NCN τ ^ CH-SCH -CH-NHCNH-R 1 H 2 having a straight or branched C 1 -C 4 alkynyl group, preferably -CH 4 = CR or - (CH 2) n C 1 -C 2 CR, wherein R is H or CH 2 and n is 1 to 6, and for the preparation of their pharmaceutically acceptable acid addition salts, characterized in that a) a compound of the formula ch3 N- ^ 5 [III VII HX CB2X or an acid addition salt thereof, wherein X is a leaving group, preferably halogen, reacting with a compound of the formula NCN HSCl 2 Cl 2 NHCNH-R 1 VIII wherein R 2 "is as defined above, or b) a compound of the formula N 2 O 2 CH 2 SCH 2 CH 2 NHR 2 29 6 9 0 6 9 or an acid addition salt thereof, is reacted with a compound of formula RgNHR1 III wherein R1 is as defined above and one of R (. and R 9 is hydrogen and another group of the formula NCN II -C-SR wherein R is a substituent such that -SR is a leaving group, and the resulting compound of formula I is optionally converted to a pharmaceutically acceptable acid addition salt. 69069 30 For the preparation of therapeutically active compounds N-cyano-N'- (2- (4-methyl-5-imidazoly) methylthio7-ethyl) -N "-alkynyl-guanidine with form CH-N- [| I NCN I TT CH-SCH -CH-NHCNH-R1 H zz A color R3, with a ring or a C, -CQ-alkynyl group, företrädes- 4. J 4 vis -CHC ^ CR eller - (CH0) C = CR, color R är H eller CH-, och n är lin 3 ch3 1-6, och farmaceutiskt godtagbara syraadditionssalter därav, kännetecknat därav, att man a) omsätter in the case of the formula CH, X 3 N-- i 11 VII ^ tr '' '- ch, x H or in the case of a sugar formulation, in which the formula X is a group consisting of halogen, in the case of the halogen NCN HSCH 2CH jNHCNH-R1 VIII color R3, anger samma som ovan, eller b) omsätter en förening med formeln ^ CH3 N-1 Us H CH2SCH2CH2NHR5