DK143701B - ANALOGY PROCEDURE FOR PREPARING SUBSTITUTED QUINOXALINES - Google Patents

Description

(19) DENMARK

| p 02) PRESENTATION SCRIPT (11) 11 + 3701 B

Dl THE RECTORATE FOR

PATENT AND TRADEMARKET SYSTEM

(21) Application No. 1 ^ 03/77 (51) | ntC | 3 C Q7 D A03 / 06 (22) Filing day 50 · Mar · 1 977 (24) Running day 50 Mar. 1977 (41) Aim. available 1. October 1977 (44) Posted Sep 28 1981 (86) International application no.

(86) International filing day (85) Continuation day - (62) Master application no. -

(30) Priority 31 Mar 1976, 672123, US Feb 23 1977, 771118, US

(71) Applicant ISTITUTO CHEMIOTERAPICO ITALIANO S.P.A., Milan, IT.

(72) Inventor Aide Garzia, IT: William Ferrari, IT: Andrea Bot = tazzi, IT.

(74) International Patent Bureau.

(54) Analogous process for preparing substituted qui = noxalins.

The present invention relates to an analogous process for the preparation of novel substituted quinoxalines of the general formula I as claimed, wherein R represents a hydrogen atom or an alkyl group having 1 to 5 carbon atoms. The quinoxalins in question are useful for the control of cholera.

Cholera is a highly contagious disease caused by an O vibrio organism. Although the infection can be spread by contact between persons, the most common source of infection is contaminated water supplies, the pollution usually originating from wastewater containing the organism. Cholera is usually combated by administering appropriate medication to the person (s) suffering from the disease. However, there are many benefits to treating

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2 143701 prophylactically exposed persons and treat the contaminated water to prevent or greatly reduce the incidence of the disease.

The invention is based on the recognition that substituted quinoxalines (hereinafter referred to as "C compounds") of the general formula I as claimed, wherein R represents a hydrogen atom or an alkyl group having 1 to 5 carbon atoms, e.g. an ethyl or propyl group, are useful for controlling cholecrosis organisms. They are especially useful for prophylactic administration before the development of the symptoms of cholera.

The C compounds concerned include, but are not limited to, CO-1: 2- [2- (2-amino-4-pyrimidinyl) ethenyl] quinoxaline-1,4-dioxide CO-2: 2- [2- (2- amino-6-methyl-4-pyrimidinyl) ethenyl] -quinoxaline-1,4-dioxide.

The process of the invention is characterized in that in a molar ratio of substantially 1: 1, quinoxaline di-N-oxide-2-carboxyaldehyde or a lower alkyl acetal thereof is reacted and a compound of the general formula II as claimed. wherein R has the aforementioned meaning, in the presence of a strong acid as a catalyst.

The reaction is advantageously carried out in a suitable solvent, e.g. a lower alkanoic acid such as formic acid or acetic acid. The reaction is carried out at reaction temperatures sufficient to cause the reaction to proceed, and these temperatures may range from ambient temperature, e.g. 0 ° C, to elevated temperatures e.g. 80 ° C or more, and is e.g. preferably from ca. 25 to 50 ° C or more. When the reaction has ended, e.g. after approx. 10 to 24 hours, the product is advantageously recovered by crystallizing it from water.

Suitable catalysts for carrying out the process according to the invention are generally known as strong acids and any known strong acid can be used. Suitable strong acids include, but are not limited to, hydrochloric, hydrobromic, sulfuric, nitric, arylsulfonic acids, e.g. toluene sulfonic acid, or trichloroacetic acid. The acids are generally used in an amount ratio of approx. 0.5 to 2 moles of acid per mole quinoxaline starting compound.

The quinoxaline di-N-oxide-2-carboxyaldehyde and its acetals used as starting material for the preparation of the compounds of the invention are known compounds. The acetals can be prepared in accordance with the method disclosed by Haddadin et al., British Patent Specification No. 1,305,128, Example XIII. The aldehyde is described in U.S. Patent No. 3,371,090.

The pyrimidine compounds used as starting materials are also known. They are commercially available and the usual commercial quality is suitable. They should preferably be of good quality, free of harmful material.

The C compounds in question are useful for controlling cholesterol-causing organisms, e.g. Vibrio-organisms. The compounds have a low toxicity and are suitable for use in oral administration for the prophylactic control of cholera. They are advantageous in that they not only confuse carbonaceous organisms, but can selectively combat the carbonate organisms without adversely affecting the desired balance of organisms in an environment, e.g. the human biological system.

The process according to the invention is further elucidated by the following examples.

Example 1

To a reaction vessel was added 15 ml of 99% formic acid, 1.15 g 96% sulfuric acid, 1.09 g (0.01 mole) of 2-amino-4-methylpyrimidine and 2.36 g (0.01 mole). quinoxaline-di-N-oxide-2-carboxyaldehyde dimethyl acetal. The mixture was heated to 45 to 50 ° C and kept at this temperature for 10 hours. It was then cooled and diluted with 35 ml of cold water and the pH adjusted to approx. 5 with sodium hydrogen carbonate. A yellow crystalline precipitate formed. It was filtered off and washed with water. 1.8 g, 64% yield, of 2- [2- (2-amino-4-pyrimidinyl) -ethenyl] -quinoxaline-1,4-dioxide, m.p. 237-239 ° C with decomposition.

The product, which was named CO-1 for convenience, was tested against five strains of vibrio cholerae at concentrations of 10, 30 and 100 micrograms per gram. ml. The results are listed in Table 1.

Tests were also performed to see if the compound was effective against vibrio cholerae El Tor Ogawa 6 in the presence of sewage. Sewage water samples were obtained from the sewer system in the city of Modena, Italy.

They were centrifuged to separate solids and the supernatant was used for the samples. The results are listed in Table 2. At 10 µg / ml of CO-1 there was no growth of 3 of the organisms after 48 hours and only border growth of the other two at 100 µg / ml.

4 143701

Compound CO-1 is administered to humans exposed to cholera under epidemic conditions at doses of 0.3 to 5 g per day. person per day for 3-5 days. None of the treated persons are infected, but numerous persons with whom they are in daily contact are infected. It is established that CO-1 is effective in oral administration as a prophylactic agent for the prevention of cholera. CO-1 is also advantageous in that it selectively controls cholera, e.g. inhibits the growth of carbonaceous microorganisms without adversely affecting the organisms of the human biological system.

Example 2

To a reaction vessel was added a solution of 1.15 g of 96% sulfuric acid in 25 ml of acetic acid, 1.23 g (0.01 mole) of 2-amino-4,6-dimethylpyrimidine and 1.9 g (0.01 mole). 2-Formyl-quinoxaline di-N-oxide. The mixture was heated to 40 ° C for 16 hours, then cooled, diluted with water and adjusted to pH 5 with sodium bicarbonate solution. The yellow crystalline precipitate formed was filtered off and washed to give 2.1 g, 71% yield, of 2- [2- (2-amino-6-methyl-4-pyrimidinyl) -ethenyl] -quinoxaline-1, 4-dioxide, which for convenience is called C0-2. It melts at 240 ° C with decomposition.

The product was tested against the 5 strains of vibrio cholerae as described in Example 1. The results are shown in Table 1. There was no growth after 48 hours of three of the organisms at 10 µg / ml of CO-2 and only borderline growth of the other two. at 100 µg / ml.

The compound CO-2 is tested for toxicity in the same manner as described for CO-1 in Example 1. Similar results are obtained which demonstrate that the compound is suitable for prophylaxis.

Compound CO-2 is administered to humans exposed to cholera under epidemic conditions at doses of 0.3 to 5 g per day. day for 3-5 days. None of the treated persons are infected, but numerous persons with whom they are in daily contact are infected. It is established that CO-2 is effective in oral administration as a prophylactic agent for the prevention of cholera.

Compounds containing 6-ethyl, 6-propyl, 6-butyl and 6-pentyl substituents, respectively, instead of the 6-methyl substituent can also be used as prophylactic agents.

5 143701

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Table 2

Sample Concentration _ Effect after__of CO-1_24 hours 48 hours 5 days

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The compound was tested for acute toxicity by various modes of administration in four animal species, namely mice, rats, guinea pigs and rabbits. The compound was found to have a low toxicity. The test results are given in Tables 3, 4.5 and 6 below.

Table 3

Acute toxicity of CO-1 in female mice

Dose Dead / treated animals by mg / kg 1 day 2 days 4 days 7 days

Endoperitoneal administration 2000 · 6/6 6/6 1000 6/6 6/6 500 6/12 250 0/18

Oesophageal administration 0 (x) 0/6 4000 1/12 1/12 2000 0/12 1000 0/12 (x) By gastric probe administration and by the carrier only.

7 143701

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Table 5

Acute toxicity of CO-1 in guinea pigs by oesophageal administration_

Dose mg / kg__Death / treated within 21 days 500 0/4 1000 1/4 2000 5/6 4000 6/6 0 (x) (x) Only carrier was administered.

Table 6

Acute toxicity of C0-1 in rabbits by oesophageal administration_

Dose Dead / Treated Body Weight in g (m + SE) mg / kg within 7 days_Start_Closing 2000 0/2 * 2250-2150 2180-2140 1000 0/4 2037-104.3 1922.5 + 71.5 0 (x) 0 / 4 2135 + 75 2262 + 215 500 0/2 2000-2100 1650-1550 (x) Only carrier was administered

There were two dead out of seven treated animals within 4 days. In view of the favorable data for acute toxicity, the compound was administered orally in sub-acute but relatively large doses to mice and rats for 15 days. Data on the effects on mortality, weight, liver and kidneys were collected. These data are given in Tables 7 and 8 below.

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Table 8

Subacute toxicity of CO-1 in female rats Daily dose: 2 g / kg / day of CO-1 by gastric probe administration for 21 days

Oral treatment Dead / Body weight ig (m + SE) __treated Start_Close_ Carrier 2/6 (x) 200.0 + 4.1 233.2 + 5.1 CO-1, 2 g / kg / day 1/6 (x) 204.1 + 2.0 210.6 + 9.6 (x) Death accidentally caused by isophagus incannalutation. This diagnosis was confirmed by the post-mortem examination.

Daily dose: 2 g / kg / day of CO-1 by gastric probe delivery for 21 days

Oral Treatment Average Percent Weight of Healthy Organs (m + SE) __Lung_Lever_Nerbs_ Carrier (3 Animals) '0.85 + 0.06 3.45 + 0.07 0.95 + 0.04 CO-1

(5 animals) 1.07 + 0.09 NS 4.54 + 0.10 HS (x) 1.04 + 03 NS

(x) Death caused by mistake of oesophagus incannalutation. This diagnosis was confirmed by the post-mortem examination. In view of the favorable subacute toxicity, the conical toxicity of female mice was investigated. The results are given in Table 9 below.

Given the favorable results in chronic toxicity, a teratogenetic study was performed with male and female mice and rats. The number of live-born cubs was comparable to that of the control animals. No malformations were observed in any of the groups. The data are listed in Table 10 below.

143701 11

Table 9

Chronic toxicity in female mice Daily treatment by gastric probe administration for 18 weeks (4.5 months) __ ^ a. Mortality and body weight ___

Oral treatment Dead / Body weight ig (m + SE) _ Treated Start _Completion Carries 3/10 28.2 + 1 33.0 + 1.1 C0-l, 500 mg / kg / day 2/10 30.4 + 0.9 30.0 + 0.7 C0.1, 250 mg / kg / day 0/10 27.3 + 0.5 26.7 + 0.7 b. Urinary excretion. Urine output secreted by 6 animals in 6 hours

Oral Treatment Urine Amount (ml)

Control 6 CO-1,500 mg / kg / day 7 C0-l, 250 mg / kg / day 6.5 c. Blood glucose. Mean values for 6 animals. Blood samples were taken 24 hours after the last dose

Oral treatment Blood glucose

Control 1.14 CO-1, 500 mg / kg / day 1.06 CO-1, 250 mg / kg / day 1.10 d. SGPT and SGOT. Mean values for 6 animals. Blood samples were taken 24 hours after the last dose

Oral treatment Units / ml

SGOT SGPT

Control 125 5 CO-1, 500 mg / kg / day 159 6 CO-1, 250 mg / kg / day 118 5 e. Fresh weight of organs

Oral treatment Guarding relationship between healthy organ and body (m + SE, 4 animals) _ Kidneys_Heart_Liver_Lunger

Control 0.938 + 0.044 0.481 + 0.055 4.57 + 0.15 0.674 + 0.044 CO-1.500 mg / kg / day 1.07 + 0.04 0.47 + 0.02 4.66 + 0.91 '1.011 + 0.110 CO-1,250 mg / kg / day 0.87 + 0.08 0.60 + 0.08 4.57 + 0.25 0.731 + 0.035 12 143701

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