ES2234438B2 - PIRIDOTIENOPIRIMIDINAS REPLACED BY GENERAL FORMULA I AS ANTIPROTOZOARS FOR AQUACULTURE AND PREPARATION PROCEDURE. - Google Patents

Abstract

The present invention relates to the process of preparing substituted pyridothienopyrimidines of general formula I and their applications in aquaculture for the treatment of scuticociliatosis in molluscs, crustaceans and fish.

Description

Substituted pyridothienopyrimidines of formula general I as antiprotozoa for aquaculture and procedure preparation.

The present invention relates to the preparation and antiprotozoal activity against the ciliated Philasterides dicentrarchi of pyrido [3 ', 2': 4,5] thieno [3,2- d ] pyrimidines of general formula I which are useful as antiprotozoals in aquaculture, and thus they are effective for the treatment of scuticociliatosis in molluscs, crustaceans and fish,

one

where the substituent R1 can be an aryl group, especially phenyl with or without substituents, or an alkoxy group (OR), especially with a carbon chain of 1 to 4 carbons, or a mono, di or trisubstituted amino group, (NH2, NRH, NRR '), open or cyclic chain, including groups piperidino, morpholino, thiomorpholino and, preferably, piperazino; where the substituent Y can be hydrogen (H) or a cyano group (CN); where the substituent X is an aromatic group, preferably a phenyl group with or without substitution; and where the R 2 substituent is a mono, di or trisubstituted amino group, (NH2, NRH, NRR '), open chain or cyclic, including piperidino, piperacino, morpholino and thiomorpholino groups and, preferably piperacino.

Among the diseases that cause greater economic damage to aquaculture, due to the ease with which pathogens are transmitted in closed circulation tanks, is scuticociliatosis, which is a disease of crustacean and mollusc fish, which affects both those raised in aquaculture facilities such as free specimens, and which is produced by ciliated parasites of the Scuticociliatida order such as Uronema marinum, Uronema nigricans, Anophryoides haemophila and Mesanophrys sp (Cheung PJ; Nigrelli RF; Ruggieri D. J. Fish. Dis 1980 , 3 , 295-303, Purple JF; Small EB Reviews in Fisheries Science 1995 , 3 , 275-354, Cawthorn RJ; Lynn DH; Despres B .; MacMillan R .; Maloney R .; Loughlin M .; Bayer R Dis. Aquat. Org . 1996 , 24 , 143-148. Munday BL; O'Donoghue PJ; Watts M .; Rough K .; Hawkesford T. Dis. Aquat. Org . 1997 , 30 , 17-25).

In recent years scuticociliatosis outbreaks caused by the protozoan Philasterides dicentrarchi have been detected causing great mortality in the farms of sea bass, denton and turbot (Dragesco A .; Dragesco J .; Cost F .; Gasc C .; Romestand B. ; Raymond J .; Bouix G. Europ. J. Protistol . 1995 , 31 , 327-340. Iglesias R .; Paramá A .; Alvarez MF; Leiro J .; Fernández J .; Sanmartín ML Dis. Aquat. Org . 2001 , 46 , 47-55. Iglesias R .; Paramá A .; Álvarez MF; Leiro J .; Sanmartín ML Dis. Aquat. Org . 2002 ) to the point of constituting one of the most important parasitosis in aquaculture, responsible for great economic losses, and that remains without a really effective treatment, having demonstrated the lack of activity of many drugs.

Detailed description and preparation of formula substances general I

In this report, by "alkoxy of C_ {1-4} "means any radical of linear or saturated branched chain containing oxygen and 1 to 4 carbon atoms, in which the radical is centered on the oxygen. Groups are included in the scope of this expression. methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, isobutoxy, t-butoxy and Similar.

For the preparation of the pyrido [3 ', 2': 4,5] thieno [3,2- d ] pyrimidines with general structure I, where the R 1 substituent may be an aryl group, especially phenyl with or without substituents , or an alkoxy group (OR) especially with a carbon chain of 1 to 4 carbons, or a mono, di or trisubstituted amino group (NH2, NRH, NRR '), open or cyclic, including piperidine groups , morpholino, thiomorpholino and, preferably, piperazino; where the substituent Y may be hydrogen (H) or a cyano group (CN); wherein the substituent X is an aromatic group, preferably a phenyl group with or without substitution; and where the R2 substituent is a mono, di or trisubstituted amino group (NH2, NRH, NRR '), open or cyclic, including piperidino, piperacino, morpholino and thiomorpholino groups and, preferably, piperacino, was based on 3-aminothieno [2,3-b] pyridin-2-carboxamides containing the selected groups X, Y and R1 (Quintela JM; Combiner C .; Veiga C .; González L .; Botana L .; Alfonso A .; Riguera R. Bioorganic Med. Chem ., 1998 , 6 , 1911-1925) and cyclized with ethyl orthoformate to give the pyrido [3 ', 2': 4,5] thieno [3, 2-d] pyrimidin-4 (3H)-corresponding areas, which were converted to 4-chloro-pyrido [3 ', 2': 4,5] thieno [3,2-d] pyrimidines (general formula I, R_ { 2} = Cl) with phosphorus pentachloride and, finally, treatment with a variety of nucleophiles results in chlorine substitution and introduction of the R2 substituent and leads to compounds I.

The method of the invention for synthesis of the pyridothienopyrimidine develops according to the following example:

Example 1 Preparation of 8-cyano-7-ethoxy-9-phenylpyrido [3 ', 2': 4,5] thieno [3,2-d] pyrimidines 4-substituted

a) A solution containing the appropriate 3-aminothiene [2,3- b ] pyridine-2-carboxamide (for example for R1 = OEt, X = Ph, Y = CN: 3-aminotiene-5-cyano- 6-ethoxy-7-phenyl- [2,3-b] pyridine-2-carboxamide), (1g, 3 mmol) in 15 mL of ethyl orthoformate containing a catalytic amount of p-TsOH, were refluxed until the product disappeared of departure. The resulting solid is filtered and the 8-cyano-7-ethoxy-9-phenylpyridido [3 ', 2': 4,5] thieno [3,2-d] pyrimidin-4 (3H) -one is used without purification Additional for the next stage.

b) A solution containing 8-cyano-7-ethoxy-9-phenylpyrid [3 ', 2': 4,5] thieno [3,2-d] pyrimidin-4 (3H) -one (2.37
mmol) obtained above, and phosphorus pentachloride (3.55 mmol) in phosphorus oxychloride (6 mL) was refluxed until the starting product disappeared. The solvent was evaporated in vacuo and the 4-chloro pyrido [3 ', 2': 4,5] thieno [3,2-d] pyrimidine contained in the solid residue was purified by silica gel chromatography.

c) A solution containing the 4-chlorine pyrido [3 ', 2': 4,5] thieno [3,2-d] pyrimidine  above (0.27 mmol) and the amine selected to introduce the R2 group, (0.32 mmol) in 10 mL of ethanol, was refluxed to disappearance of the starting product. The solvent was evaporated at empty and the 8-cyano-7-ethoxy-9-phenylpyridido [3 ', 2': 4,5] thieno [3,2-d] pyrimidine Resulting 4-substituted was purified by chromatography or crystallization.

Antiprotozoal activity

The compounds object of the invention are active against the protozoan Philasterides dicentrarchi , which causes scutiliacytosis in fish, molluscs and crustaceans. The activity was measured in the ciliates obtained from the natural cavity, maintained in cultivation conditions that ensure their capacity for infection of turbot, sea bass and other species of interest in aquaculture. The activity tests were carried out in DMSO / PBS and in DMSO / seawater, and are expressed in DL (Lethal Dose): minimum concentration (mg / L) necessary for the removal of 99% of the ciliates.

The obtained DLs indicate that the pyridothieno [3,2- d ] pyrimidines I indicated here, and especially those containing at least one piperacino group as a substituent of the heterocycle, show great antiprotozoal activity against the ciliated Philasterides dicentrarchi . Specific example of some of these new parasitic agents is pyridothienopyrimidine 1 (general formula I, R1 = OEt, X = phenyl, Y = CN and R2 = piperazine) (DL = 12.5 mg / L) .

The histamine release inhibitory action (Quintela JM; Peinador C .; Botana L .; Estévez M .; Riguera R. Bioorganic Med. Chem ., 1997 , 5 , 1543-1533. Quintela JM; Peinador C .; Veiga C. ; González L .; Botana L .; Alfonso A .; Riguera R. Bioorganic Med. Chem ., 1998 , 6 , 1911-1925. Quintela JM; Peinador C .; Veiga C .; Botana L .; J Alfonso A .; Riguera R. Eur. J. Med. Chem ., 1998 , 33 , 887-897) and anti-inflammatory activity (Quintela JM; Peinador C .; González L .; Riguera R .; Rioja I .; Terencio MC; Ubeda A. ; Alcaraz MJ J. Med. Chem ., 1999 , 42 , 4720-4724) of some of these compounds has been described by the authors of this invention previously.

Also, compounds that possess the piperacino group linked to various heterocycles such as benzoisoquinolines and nitroimidazoles are known to possess antiprotozoal activity and have been subject to prior patents (Wade, P .; Vogt Berthold R .; US Patent 4,070,465, 1978. Makovec, F. .; Senin, P.; Rovati, L .; Brit.UK Pat. Appl. GB 79-1661719790514, 1979 ).

Claims (1)

1. Use of Pyrido [3 ', 2': 4,5] thieno [3,2-d] pyrimidines substituted of general formula I 2 where the substituent R1 can be an aryl group, especially phenyl with or without substituents, or an alkoxy group (OR) especially with a carbon chain of 1 to 4 carbons, or a mono, di or trisubstituted amino group, (NH2, NRH, NRR '), open chain or cyclic, including piperidino groups, morpholino, thiomorpholino, and preferably piperazino; where he substituent Y may be hydrogen (H) or a cyano group (CN); where the substituent X is an aromatic group, preferably a group phenyl with or without substitution; and where the substituent R2 is a mono, di or trisubstituted amino group, (NH2, NRH, NRR '), of open or cyclic chain, including piperidino groups, piperacino, morpholino and thiomorpholino and, preferably, piperacino, alone or in combination with other agents, in the preparation of any kind of pharmaceutical form, food additives or in feed processing for disease treatment protozoan in fish, crustacean and mollusc culture and activities of aquaculture.