DE1816086C3 - 2-Aminoalkyl-2H- (1) -benzothiopyrano (4,3,2-cd) indazole compounds and process for their preparation - Google Patents

Description

The invention relates to 2 aminoalkyl-2H- (1) benzothiopyrano (4,3,2-cd) indazole compounds, their acid addition salt and process for their preparation. The compounds according to the invention have the formula

N-N --- Λ - Ν

\ Ίί ι "
■ s -.

R,

R,

wherein A is an alkylene radical having 2 to 4 carbon atoms. Q is hydrogen or halogen, Ri and R2, which can be identical or different, are an alkyl group with 1 to 4 carbon atoms, which with the nitrogen atom to form a heterocyclic ring with 5 to 7 Ring members can be linked, and W the aldehyde group or a methyl or hydroxymethyl group means, or their acid addition salts.

According to one embodiment of the method are erfindungsgemäen lndazolverbindungen that ^the: Ormel

Ν — Ν — Α — Ν

(Ia)

CH,

have prepared by reacting a haloalkylamine the formula

XAN (Ri) R ₃
with an indazole of the formula

N NH

A λ A

ι if ^ l I

(U)

St. ₁

in the presence of a base. The product is isolated as the free base or, if appropriate, as an acid addition salt, obtained by reacting the free base with an acid. The reaction conditions can be varied widely and are not critical. In particular, the ratio of the reactants can be varied as desired and, for example, equimolar amounts or an excess of one or the other reactant can be used. The haloalkylamine is preferably used in excess. One of the various alkali and alkaline earth metal hydrides and amides can be used as the base present in the reaction. A preferred base is sodium hydride. The reaction is usually carried out in an organic solvent. In general, an inert aprotic solvent such as an aliphatic or aromatic hydrocarbon, dimethylformamide, dimethyl sulfoxide or saturated ether is used. A preferred solvent for the reaction is xylene. The reaction is carried ^r t at temperatures within a range of ctv ". 20 to 140 "C and is complete within about 4 and 20 hours.

According to another embodiment of the reaction, indazole products of the above formula Ia are prepared by using dialkylaminoalkylhydrazine

H ₂ N-NH-AN (Ri) R ₂
with a haloxanthene compound of the formula

O X

(Uli

ClI,

where A, Q, Ri and R. have the above meaning, implements. The product is obtained as a free base or, in any case, as sikiresal /. By reaction the free base with an acid, isolated. The reaction conditions can be varied widely are and are not critical. In particular, the ratio of reactants can be varied, by using either equimolar amounts or an excess of one or the other of the reactants The dialkylaminoalkylhydrazine is preferably used in excess. That used Solvent must be inert in the reaction.

Pyridine, phenol, a lower alkanol such as methanol or an excess of are suitable, among others Dialkylaminoalkyl hydrazine. Pyridine is preferred as the solvent. The reaction becomes ordinary carried out at temperatures above about 75 ° C., preferably at about 95 to 140 ° C. The reaction at these high temperatures takes about 16 to 24 hours to complete. In order to isolate the Product from the reaction medium, the latter is made alkaline and then in one of the following Further treated in a manner: For example, the reaction medium can be subjected to steam distillation, to separate the volatile components and then remove the product from the residue with a solvent, like ether, extract. You can also take up the residue in glacial acetic acid and the solution with water dilute, filter and extract the alkalized filtrate with chloroform, so that the product in in dissolved form. Another preferred isolation method is that the reaction mixture extracted with a solvent such as benzene and the product from the extracts on chromatographic Paths isolated.

According to a further embodiment of the production process for the invention Compounds can be the indazoles of the above formula I, in which W is an aldehyde group (—CHO) or is a hydroxymethyl group (CH2OH), by adding a compound of the formula Ia of Subjected to microbiological conversion. For this purpose, the starting indazole is used together with a Microorganism iii a suitable growth medium introduced under fermentation conditions which are sufficient to convert the starting indazole into the corresponding Indazole of formula 1, wherein W is an aldehyde group (-CHO) or a methanol group (-CH2OH) is converted. For fermentation the microorganism Aspergillus sclerotiorum (Imperial Mycological Institute No. 56 673) can be used will.

According to the invention, the microorganisms are cultivated in an aqueous nutrient medium. The composition of the medium is based on general considerations in fermentation technology, which are known per se; the medium contains one source for organic carbon and another for organic nitrogen. Optionally, a source of inorganic nitrogen and additional Growth factors, mineral salts and trace substances must be present. As sources of organic carbon serve for example starch, corn flour, sugar oiler glycerin. The organic nitrogen can be provided are obtained from any source such as casein, soybean meal, almond nut meal, cottonseed meal, Wci / cnklecr, barley or oat flakes, lactalbumin, enzymatically processed casein, tryptone and flour peptone. A suitable source of inorganic Stickstolf is ammonium chloride or ammonium sulfa. Growth factors can be provided

through soluble distillery waste, yeast autolysate, yeast extract or residues from molasses fermentation. Suitable mineral salts include sodium chloride, Potassium sulfate and magnesium sulfate. Examples of trace minerals are copper, cobalt, manganese, iron and zinc. An antifoam agent can be used to control foam formation during fermentation such as polyglycol, silicone, lard oil, mineral oil, vegetable oil or an anti-foam agent isi form one higher alcohol.

The fermentation is expediently carried out in various successive stages: 1. Establishing inclined cultures, 2. sowing stage, 3. preliminary stage for conversion and 4th conversion stage. In the first three stages, the microorganism and the fermentation medium are produced in sufficient quantities. At the conversion stage, the starting indazoline is introduced into the inoculated growth medium, and the fermentation is carried out until the substrate has been converted into the desired end product by microbiological conversion. The fermentation conditions, such as pH value, temperature, production of the end product, etc., can be quite different. The initial pH value of the medium is expediently between 5.5 and 7.5, the optimum being a pH value of 6.4. Optionally, a buffer such as calcium carbonate can be incorporated into the medium. The initial concentration of the indazole substrate in the fermentation medium can vary between about 0.1 and 5 mg / ml or higher; the optimum is around 0.5 mg / ml. The indazole can be incorporated into the fermentation medium in any number of ways, e.g. B. as a free base, as a methanol solution of the free base, or preferably as an aqueous solution of a salt of the free base. The temperature can be between 20 and 40 ^° C., preferably it is 29 to 30 ^° C., at which temperature the reaction takes place in about 40 to 44 h. During the fermentation process, the Gc is mixed and aerated. The air supply is preferably at a speed of about I ¹ /? Volume of air / volume of fermentation medium held in the minute. Stirring is effected by the usual means, e.g. B. with a propeller agitator, which works during the preliminary stage of the conversion at a speed of about 200 rev / min and during the implementation at about 250 rev / min. The speed of the reaction can be followed, as usual, by taking samples and examining the samples during the reaction. The samples taken are extracted and the extracts analyzed by thin layer chromatography. To isolate the product, the fermentation mixture is filtered and the filtrate and the mycelium cake are extracted separately with an organic solvent such as methylene chloride. The products are extracted in pure form from the extracts. The extracts are preferably concentrated, redissolved in ßcn / .ol and chromatographed on a column, e.g. B. an alumina column, fractionated. Preferred performance or eluent means for obtaining the products from the column are benzene or ethyl acetate or mixtures of the two. When eluting the column with benzene, e.g. 1?. with individual portions of benzene containing successively increasing proportions of ethyl acetate, one obtains first the 5-carboxyaldchydindazole and then the corresponding 5-methanol product.

According to a further implementation form of the Process according to the invention, indazoles of the formula 1. in which W is a melano group (—CH.OH). prepared by adding a compound of formula J. in which W is an aldehyde group (-CHO), reduced with an alkali borohydride. The response will be at Carried out at normal temperature and sodium borohydride is preferably used.

The compounds according to the invention present in the form of free bases form by reacting

tion with organic and inorganic acids addition salts. Examples of such acid addition salts are the salts of inorganic acids, such as the hydrochloride, the hydrobromide, the hydroiodide, the Sulfate, the carbonate and the phosphate and salts

is organic acids such as succinate, benzoate, acetate, Citrate, malate, maleate, p-toluenesulfonate, gluconate, Benzenesulfonate. and the sulfamate and salts with dibasic acids, such as methylene bis (hydroxynaphthoic acid). The acid addition salts are

moderately formed by mixing the free base with at least the equivalent amount of the acid in a solvent in which the salt, especially in the cold, is sparingly soluble, so that it is easily as a solid Phase can be obtained. It is claimed here

2s the protection for salts of the compounds according to the invention in general, so that the choice of anion is not important, since the cation is pharmacological The active part. The selection and Providing salts for purposes of the invention

must be left to the expert, who will also depend on the application. Salts, for a specific purpose, e.g. B. for types of application in which toxicity plays a role, not seem suitable, valuable inter-

represent chemical products that can be easily converted into non-toxic salts. Compared to the free acids, which are used for many purposes , their salts, in particular with inorganic acids, are preferred if higher water solubility is desired.

The compounds according to the invention are valuable medicaments. In particular, they have a high grade antibacterial effectiveness and can therefore be used as antibacterial agents for local application or for

Use 4s oral or parenteral administration. In In vitro, the compounds have a killing effect even in doses as low as 20 y / ml, which is due to the usual standard tests against organisms such as S. pyogenes and E. coil was determined. the

so compounds, especially those according to formula I, where Q stands for a chlorine atom, have a noticeable schistosomicidaic effectiveness. Such products are therefore valuable as a means of combating the Schistosomiasis when administered orally or parenterally

ss ehung. Such connections have good Effective wedge on when they are mice in standard experiments by means of the gastric conduit or in the diet food at doses of about 200 mg / kg and 0.125%, respectively. The compounds according to the invention have

fto also an antitrichomonalc effectiveness on and are therefore applicable as antitrichomonal agents for local application. In vitro, for example, were found the products as trichomonicid in doses of about 25} '/ ml.

6s The attempts that lead to the above Results were carried out as follows: Kupferberg's medium (containing 250 μg / ml Nairium penicillin G and slreptomycin sulfate), which contains

L'iner sufficient Λη / iihl organisms of one 24-Suinden-Kupfcrbergkulmr had become legged. so that 10,000 Tiichomonads / ml was obtained at 37 C incubated with an aqueous solution or suspension of the compound to be investigated. After 48 minutes Incubation was the number of viable trichomonads by microscopic examination definitely.

To determine the toxicity of the inventive Compounds were some representative compounds in mice at a much higher rate Dose than the effective dose administered. At these doses tested, it was found that none of the Experimental animals died. The respective amounts administered and the mode of administration are based on the following Table shows:

Connection (I) Q W. C2lh R: Mode of administration crowd Λ Il CHi CMI, CMI, (mg / kg) C ₂ Hi Cl CHi CMI, intramuscular 200 C2H4 food 379 Isopropyl Nasogastric tube 400 Cl CHi CHi intramuscular 400 C2Ü4 Pyrrolidinyl food 218 Cl CHi Pipcridinyl Nasogastric tube 200 CiHt, Cl CHi food 55 CIH H CMI, food 180 H CHjOII CjH-, Nasogastric tube 200 CiMi CMI, food 143 Cl CHO CMi-, CjH-, i. m. (hamster) 200 (JiUa Cl CH: OH CjH-, Nasogastric tube 100 CM-U Nasogastric tube 100 1. m. 400

example 1

To a suspension of sodium hydride (3.6 g) in a 50% dispersion of oil and 100 ml of xylene is added

5-methyl-2H- [1] benzothiopyrano [4.3,2-cd] indazole
(12.9 g) were added. The mixture is refluxed for 2 hours and then a solution of 2-chlorotriethylamine (6.8 g) in 100 ml of xylene is added and the mixture is refluxed for 20 hours. Water is added to the cooled reaction mixture and the organic layer is drawn off, which is then dried over anhydrous magnesium sulfate. The mixture is filtered and evaporated iind gives the desired product, 2- [2- (diethylamine

dazol; Mp. 54 to 56 ⁰ C, after recrystallisation from 2,2,4-trimethylpentane.

Example 2

(a) A solution of 3-dimethylaminopropyl hydrazine (59 g. J. Med. Chem. 7, 493). 100 ml of pyridine and 130 g of a 60:40 mixture of i-chloro-4-methyl- and 4-chloro-1-methylthioxanthen-9-one (J. Am. Chem. Soc, 74, 4296) is under for 18 hours Held at reflux and cooled. Potassium hydroxide is added to the resulting mixture and subjected to steam distillation to remove volatiles. The remaining product is nitrated and the filter cake is dissolved in glacial acetic acid. The solution thus obtained is treated with water, filtered and the filtrate made alkaline with sodium hydroxide and extracted with chloroform. The chloroform extracts are dried over anhydrous potassium carbonate and filtered and the filtrate is concentrated to an oil which is taken up in ether. The ethereal solution is treated with anhydrous hydrogen chloride and the resulting product, which separates out, is separated. The product is 2- [3-Di-

methylamino) propyl] -5-methyl-2H- [1] benzothiopyrano [4,3 ^ -cd] indazole dihydrochloride 3/4 hydrate; Mp. 200 to 204 C.

To convert the salt into another salt of the

The same free base can be worked as follows: A solution of the dihydrochloride (3.96 g) in 15 ml of water is slowly added to a cooled solution of benzylpenicillin potassium (7.44 g) in 15 ml of water while stirring. The benzyipenicillin salt of the free base separates out in the form of a yellow precipitate and is filtered off and dried under vacuum.

(b) A solution of 26 g of 2-diethylaminoethylhydrazine (|. Med. Chem., Op. Cit.), 29 g of an 80:20 mixture of lo-dichloro-methyl- and 4,6-dichloro-1-methylthioxane 9-one (J. Am. Chem. Soc. Loc. Cit.) And 17 ml of pyridine is as in the previous paragraph (a) implemented and subjected to steam distillation. The residue from the steam distillation is extracted with ether and the extracts dried over anhydrous potassium carbonate and filtered. Then adds anhydrous hydrogen chloride is added and the precipitate which separates out is filtered off and dissolved in water and treats the solution with sodium hydroxide. The alkaline mixture is extracted with ether which

so extracts are dried and treated with anhydrous hydrogen chloride. 8-Chloro-2- [2- (diethylamino) ethyl] -5-methyl-2H- [1] benzothiopyrano- [4,3,2-cd] in dazene monohydrochloride hemihydrate is obtained which, after recrystallization from 2-propanol, gives a having mp. 199-202 ⁰ C.

Example 3

A solution of 2- (isopropylmethylamino) ethyl hydrazine (10 g, J. Med. Chem, op. Cit.), 50 ml of pyridine and 24 g of an 80:20 mixture of l, 6-dichloro-4-methyl and 4,6-dichloro-1-methylthioxanthen-9-one (J. Am. Chem. Soc. loc. cit.) is refluxed for 24 hours held and cooled to room temperature. After adding aqueous sodium hydroxide solution, the reaction mixture extracted with benzene and the extracts The residue is taken up in benzene and the benzene solution is chromatographed fractionated by adsorption on a clay column.

709 612/89

The column is eluted successively with benzene and then with benzene-ethyl acetate mixtures become increasingly richer in ethyl acetate. The solution eluates are concentrated and then give as The residue is the desired product, the 8-chloro-2- [2- (isopropylmethylamino) ethyl] -5-methyl-2-H- [l] benzothiopyranof4,3,2-cd] inda / .ol; up to 8HC. One obtains in the same way if one of the corresponding amounts of the respective starting materials exclude the following connections:

8-chloro-2- [3- (1-pyrrolidinyl) propyl] -5-methyl-2ll- [1] -bcnzothiopyrano [4,3,2-ed] indazole; M.p. 8b to 87 ° C. 8-chloro-2- (3-piperidinopropyl) -5-methyl-2H [1] ben / othiopyrano [4.3.2-cd] indazole; Mp 87-88 ° C. 8-chloro-2- (3-dimethylamino-2methylpropyl) -5-methy! - 2H- [l] benzothiopyrano [4,3,2-cd] indazole and

8-Broni-2- [3- (butylmethylamino) propy I] -V methyl-2H [I] -benzothiopyrano [4.3,2-ed] indazole.

In a similar way, the 8-chloro-2- [2- (2,6-dimethylpipcridinoJäthyljS-methyl ^ H-fljbenxotliiopyrano [4,3.2-cd] indazole and the corresponding substituted in the 2-position by the 3- (hexameihylenimino) propyl group Indazole can be produced, one of the appropriately substituted alkyihydrazine starting material goes out. The starting materials can be produced in the following ways:

For the production of the required dimethylpiperidinoethylhydrazine is added to a solution of 100 g of 85% technical grade hydrazine and 100 ml of water Room temperature 87.8 g of 2- (2,6-dimethylpiperidino) -äthylch'orid to, the temperature below Holds 30 ° C. When everything has been added, another 35 g of anhydrous potassium carbonate are added and the mixture Maintained under reflux for 7 hours. After adding 187 g of solid sodium hydroxide, which is made in portions adds, the resulting alkaline mixture is extracted with two portions of ether of 250 ml each and the Ether extracts dried over solid potassium hydroxide. The extracts are concentrated and taken from the residue the desired 2- (2,6-dimethylpiperidino) ethylhydrazine is obtained after vacuum distillation.

component

Glucose monohydrate
s soybean meal

Here yeast (USI ', Yeast Products Co.) sodium chloride
Dikiiliumhydrogcnphosphai (anhydrous)

2.0% 2.0% 0.5% 0.5% 0.5%

The whole thing is made with 10 N sodium hydroxide solution pH adjusted to b.4 and sterilized by heating to 121 ° C. Then tap water on 100% filled.

The growth medium mixture obtained in this way is stirred for 3 hours at 200 revolutions per minute and kept at M C so that the seeds are obtained for the first time.

Preliminary stage of conversion

Added seed from the 1st stage in an amount of 800 ml w ill / u 16 I growth medium of the composition described above, and the mixture Munden 32 _In a 30-LITCR-Fermcntator at 30 C for ^rj.

Example 4

Production of the compounds according to the invention by oxidative fermentation of the starting materials Conversion stage

Now a solution of 2- [2-Diaihylamino) ethyl] -5-methyl-2H-tl] benzothiopyrano [4,3.2-cd] indaz.ol (8 g) in 200 ml of methanol are added and another 47 hours at 30 ° C. with 250 Rev./mm stirred. The resulting fermentation medium is filtered, whereupon both the filtrate and the Rillerkuchen extracted separately with methylene chloride will. The extracts are combined and concentrated 3 :, and the residue is taken up in benzene and fractionated by chromatography over a column. A column of 5 × 125 cm is used, filled with 1 kg of powdered aluminum oxide (Chromaograph type F-20. Alcoa). 4 "Solvent fractions (about 1 liter each) are passed through the column The following composition is sent through from retnem benzene via benzene / ethyl acetate to pure Atnylacetate range:

Preparation of sloping cultures

(a) A suspension of a lyophilized culture of Aspergillus sclerotiorum (from the Commonwealth Mycological Institute, referred to as CMI 56 673, also known as IMI 56 673) in sterile distilled water is placed on a tilted agar medium containing the Pacreas digestion product of Casein, pulpy digestive product made from soybean meal and sodium chloride. After incubation at 28 ° C. for at least 1 week, 10 ml of sterile aqueous 0.1% sodium heptadecyl sulfate solution is added to four selected cultures so that the spores and vegetation are detached. The resulting mixtures are added to the growth medium described below

Preparation of "seeds" (sowing stage) The mixtures obtained from the inclined culture 65 are placed in a 30 liter fermentation jar containing 121 sterile growth medium of the following composition:

Factions

1 to 11
12 to 14
15 to 20
until 23
to 63
to 74
to 94
to 128

benzene

100%
98%
95%
90%
80%
60%
20%
0%

Ethyl acetate

0%

2%

5% 10% 20% 40% 80% 100%

this"

S.

86 °? ^:
The

fif ^{rte product is} 2- [2- (diethyIamino) -

^{emem single Ar} beitsgang was """""lish in fractions 31 to ^ ^Ukt holding fractions T if ^{Connectivity"} "isolate g ™ mp 83 to""^'^ fcieren from -Trimethylpentan..

^Product < ^which essentially ^F ' ^actions 82 to 128 occurred) is

- ^ »ethanol; Mp. 85 to 86 ° C after and recrystallization from cyclohexane.

For the purpose of working up, the presence of the desired in the operation just described Products in a given sample are conveniently determined by thin layer chromatography.

(b) A solution of the 5-carboxyaldehyde product from operation (a) in an amount of 1 g in 175 ml of Metlvinol is treated with 0.2 g of sodium borohydride. After stirring for 5 hours at room temperature, the mixture is diluted with 200 ml of ice water. The reduction product, the 2 [2- (diethylamino) ethyl] -2lf- [1] benzothiopyrano [4,3,2-cd] indazole-5-methanol, precipitates out as a precipitate and is filtered off. According to an alternative procedure, you can also concentrate the methylene chloride extracts from the fermentation mixture of seeds / (a) (which contain both 5-carboxyaldehyde and 5-meihanol) and dissolve the residue in 175 ml of methanol. The solution is then treated with sodium borohydride as above, so that only 5-methanol is obtained.

(c) If one proceeds exactly according to the method described in part (a) above, it is assumed, however, that it is water-based gen solution of 8-chloro-2- [2- (diethylamino) ethyl] -5-methyl-2H- [l] benzothiopyrano [4,3,2-cd] indazole mono- hydrochloride hemihydrate, the two products formed are 8-chloro-2- [2- (diethylamino) ethyl] -2H- [benzoihiopyranoJ ^ .S ^ -cdJindazole-S-carboxyaldehyde ιυ (m.p. 1 Jl to 132 ° C from isooctane / ether) and 8-Ch! or-2 [2- (diethylamino) ethyl] -2H- [1] benzothiopyrano [4,3,2-cd] indazole-5-meihanol; M.p. 157-158 ° C. Analogously, if you look at a methanol solution of 8-Chk; r-2- [2- (isopropylmethylamino) ethyl] - or

is 8-Chloro-2- (3-piperidinopropyl) -5-methyl-2H- [1] benzothiopyrano [4,3,2 cd] indazole goes out, the corresponding 5-carboxyaldehyde or the corresponding 5-methanol.

Claims (7)

Patent claims: 1. 2-Aminoalkyl-2H- (1) benzothiopyrano (4,3,2-cd) indazole compounds of formula I. Ν — Ν — Α — Ν wherein A is an alkylene radical having 2 to 4 carbon atoms, Q is hydrogen or halogen, Ri and R2, which can be identical or different, are an alkyl group with 1 to 4 carbon atoms, which with the nitrogen atom to form a heterocyclic ring with 5 up to 7 ring members can be linked, and W is the aldehyde group or a methyl or hydroxymethyl group means, or their acid addition salts. 2. 8-Chloro-2- [2- (diethylamino) ethyl] -5-methyl-2H- [1] -benzothiopyrano [4,3,2-cd] indazole. 3. 8-Chloro-2- [2- (diethylamino) ethyl] -2H- [1] benzothiopyrano [4,3,2-cd] indazole-5-methanol. 4. 8-Chloro-2- (3-piperidinopropyl) -5-methyl-2H- [1] benzothiopyrano [4,3,2-cd] indazole. 5. Process for the preparation of the compounds according to claim 1, wherein W is methyl, thereby characterized in that a haloalkylamine of the formula is used in a manner known per se XAN (Ri) R2
with an indazole of the formula N-NH CH, reacted in the presence of a base and the product isolated as a free base or as an acid addition salt or a dialkylaminoalkylhydrazine in a manner known per se H ₂ N-MH-AN (Ri) R ₂
with a haloxanthene compound of the formula O X cn, converts and the product as a free base or as Acid addition sal / isolated. 6. A process for the preparation of compounds according to claim 1, wherein W is -CHO or —CH2OH, characterized in that a compound of the general formula R. Ν — Ν — Α — Ν ί ί ^! j CH, oxidative fermentation by treatment with Aspergillus sclerotiorum (Imperial Mycological Institute No. 56 673). 7. A process for the preparation of compounds according to claim!, Wherein W is a methanol group is, characterized in that an indazole of the formula N- Λ -Ν R, • s - CHO reduced in a manner known per se by means of an alkali borohydride and the product as a free base or Acid addition salt isolated.