CN1031052C - Method for treatment and prophylaxis of pneumocystis cariniipneumonia and other diseases and compounds and formulations for use in said - Google Patents

Abstract

This invention relates to a method for treating giardiasis with an aromatic diamidine compound.

Description

The method of treatment and prevention pneumocystis carinii pneumonia and other disease and used compound and prescription

The present invention relates to the method for pneumocystis carinii pneumonia, be particularly related to and have the compound and the formula of medicine that contains this compounds of Pneumocystis carinii pharmaceutical activity extremely, and the method for treatment and prevention pneumocystis carinii pneumonia and treatment leishmaniasis, the method for giardiasis and malaria.

The hydrochloride of five amidines is at first discoveries (seeing United States Patent (USP) 2277861) such as Ewins, produced water-soluble salt subsequently again, see No. 2410796 United States Patent (USP)s of application such as Newberry, this patent relates to the hydroxyethanesulfonic acid salt and the hydroxypropanesulfonic acid salt of this water-soluble salt, particularly five amidines.Preceding a kind of compound is commonly referred to as hydroxyethylsulfonic acid five amidines.Pneumocystis carinii pneumonia

Hydroxyethylsulfonic acid five amidines are produced by the LyphoMed corporate system now and sell, and its trade mark is Pentam, is used for vein and intramuscular injection, the pneumonia that treatment is caused by Pneumocystis carinii, and this disease is commonly referred to as " PCP ".Because PCP patient significantly increases, the importance of hydroxyethylsulfonic acid five amidines progressively rises dramatically in recent years.It is because the unfortunate consequence that acquired immunodeficiency syndrome (AIDS) patient increases that patient increases.The PCP that estimated now almost 70% AIDS patient infection.Because the high incidence of PCP among the AIDS VICTIMS is found that hydroxyethylsulfonic acid five amidines are not only useful to treatment PCP, but also be can be used to prevent PCP, to prevent or to delay particularly first or the recurrence of AIDS VICTIMS PCP.

Yet unfortunately the side effect of hydroxyethylsulfonic acid five amidines is that it is toxic.In with the patient of hydroxyethylsulfonic acid five amidines, owing to serious ypotension, hypoglycemia and irregular pulse cause some deaths by muscle and intravenous injection treatment.Because overcome the toxicity of hydroxyethylsulfonic acid five amidines, thus the substitute of a kind of hydroxyethylsulfonic acid five amidines be badly in need of, to avoid or to reduce the adverse side effect that uses five amidines and cause.

Giardia lamblia

In the U.S., Lan Shi giardia lamblia (Giardia lamblia) is that a kind of modal intestinal parasite (is seen Peaiatr.Clin.North AM, Jun.1988,35 (3): 565-77).It is said that it is the most general in the world pathogenic enteron aisle protozoon and the important pathogenesis of gastrointestinal disorder.It is the problem of an especially severe of third world countries, and after children are infected by it stubborn problem very especially.

In the research that portion infects the Jamaica malnourished children, in those infected cases, determined a kind of paathogenic factor, the Lan Shi giardia lamblia be the most general enteropathogen (J.Trop.Med.Hyg.91 (4): 173-80, Aug.1988).Another to the research that inhabits The Nile, Egypt delta area family in, investigated 724 children, wherein only have children be shown as under study for action Lan Shi giardia lamblia feminine gender (Am.J.Epidemiol.127 (6): 1272-81, Jun.1988).

Unfortunately, although the Lan Shi giardia lamblia is a ubiquitous pathogenic protozoon like this, they are lodged in the top of small intestine, cause acute and chronic diarrhoea and nutrient malabsorption, and the medicament that is used for the treatment of now are extremely not satisfied.In fact, the treatment medicine that general choice is now used all is that main the manufacturing is used for treating other transmissible diseases, finds again afterwards that they were effective to the Lan Shi giardia lamblia.The typical medicaments of treatment giardiasis has Metronidazole plain BP.98 99, his imidazoles, Quinacrime or Nifurazolidone, but such treatment all has typical adverse side effect, and always ineffective.

Although five amidines are decades ago by known to the people, before this and do not know that it can be used for treating giardiasis.

Because infection by Giardia lamblia is in worldwide seriousness, and lacks effective medicine of treat this disease, so self-evident, extremely need a kind of more efficientlyly, the medicament of the anti-giardiasis of good therapeutic property is arranged.

Leishmania

Known leishmania is a protozoan parasite in a kind of born of the same parents, and it can cause people's severe infections.This biology " is bitten " by infected sand fly and is propagated, and invades in the reticuloendothelial system (RES).This parasite has very high growth and reproductivity in the tissue that the vertebrates host is responsible for reacting with the intrusion organism.Can expect that leishmanial this parasitic point has brought difficulty for the effective chemical therapy, and has the interaction of high complexity between parasite and host's cellullar immunologic response.In reticuloendothelial system, parasite is in the scavenger cell of existence the host in the part of its breeding cycle at least.Fusion has obviously taken place in the secondary lysosome of host cell and parasitic vacuole, and fails to prevent leishmanial breeding subsequently.Thisly be fused to parasite and obtain nutrition approach is provided, but also make parasite be exposed to host's antibody and lysosomal enzyme.

In human body, the result that Leishmania donovani (Leishmania donovani) is successfully invaded spleen and liver is exactly dead usually.Cicatrix of skin may be that crithidia cunninghami (Leishmania tropica) and the similar of it are had a liking for the skin organism (as leishmania aethiopica (Leishmania aethiopica), leishmania mexicana (L.mexican), leishmania peruviana (L.Peruviana) and Guyana leishmania (L.Guyanensis)) unique performance of infecting.It is moderate infection that leishmania brasiliensis is invaded mucocutaneous tissue.Unfortunately, only find that in screening a few relatively medicine demonstrates enough leishmanicidal activity, be worth carrying out human trial just still less.The main antimonial that relies on of treatment, this medicament also can produce the side effect of serious poisoning, particularly in those underfed patients.A kind of antimonial that is widely used in treatment is the N-methyl glucoside amine salt of metaantimmonic acid, is commonly referred to the meglumin stibnate.The toxicity of this class medicine may influence liver (hepatitis), kidney (ephritis) or heart (myocarditis).In these toxic effects, myocarditis be maximum also be the most general problem.

Initial find that five amidines are effectively when the treatment trypanosomiasis, as above selected recently, find that five amidines are very useful for the pneumocystis carinii pneumonia, particularly to those because acquired immunodeficiency syndrome (AIDS) and ill patient.Up to now, only know five amidines, rather than its congener, in the treatment leishmaniasis, some limited effects are arranged.

Since lack the medicine of the satisfaction of treatment leishmaniasis, therefore need be a kind of more efficiently, have the anti-leishmaniasis medicine of good therapeutic property.

Malaria

Human malaria is caused by the plasmodium Parasites.It is by having inhaled the parasitic mosquitoes spread of sexual body in the blood.Parasitic exotospore is grown up in mosquito, and propagates in new host's individuality by biting of mosquito.Main human pathogen is plasmodium falciparum (Plasmodium falciparum).

Malaria is one of most important health problem of tropical under-developed country.Estimate that the whole world has more than ten hundred million people to live in the malaria transmission area.Though chloroquine is a kind of effective medicine, this medicine has some side effect, the more important thing is that malarial parasite has obtained the resistance to chloroquine.

Therefore along with the appearance of chloroquine resistant strain in the malarial parasite, and the reduction renderd a service of DDT one class persistent pesticide, the malaria of torrid areas has become a kind of serious day by day problem.Malaria is the seriousness that maximum in the world this fact of transmissible disease also reflects this problem.In 1,000,000,000 populations of living in the malaria transmission area, any one given interior nearly 25,000,000 to 200,000,000 people of time suffer from malaria.

In Africa, estimate at a million people every year and die from malaria, mainly be children less than five years old.Even those were infectd under people's survival of malaria in childhood, the grownup of significant proportion still is easy to infect malaria, shows periodic parasitemia, kills plasmodium antibody although contain " protectiveness " in their serum.It is believed that in African endemy hotspot nearly all resident suffers from the mild to moderate pathogenic falciparum infection of successive throughout one's life.

Because increasing malaria pathogenic strain has resistance to main anti-malarial medicine, the problem of malaria infection becomes even more serious.More and more to chloroquine have drug-fast plasmodium falciparum product tie up in and South America, Africa and South East Asia occur.

Researchist's synthetic various chloroquines come and the struggle of new plasmodium resistant strain.But these strains have resistance to new drug.Released a kind of new drug mefloquine recently, yet its resistive new lines has also been occurred.The brand-new medicine that needs a kind of and chloroquine to have different chemical character is contained the increased popularity of plasmodium resistant strain.

The appearance of five amidines has decades, shows at first to can be used for treating trypanosomiasis.As mentioned above, have been found that five amidines are very useful to the pneumocystis carinii pneumonia, particularly to those because of suffering from the patient of acquired immunodeficiency syndrome (AIDS) immunizing power sustain damage.Equally, known five amidines rather than its congener only utilize in the treatment malaria to some extent.

Because the infectious in the world seriousness of malaria disease and lacking its effective medicine for the treatment of is self-evident, be badly in need of a kind of more efficientlyly, have the antiplasmodial product of good therapeutic property.

Pneumocystis carinii pneumonia

According to an aspect of the present invention, find unexpectedly that pneumocystis carinii pneumonia can effectively be treated with the compound that defines as structural formula I: Wherein, each R ₁All be H, perhaps two R on same amidino groups ₁Common representative-(CH ₂) _m-, m=2 wherein, 3 or 4, R ₂Be H, OCH ₃, NO ₂Or NH ₂, R ₃Be H, CH ₃Or CH ₂CH ₃N=2,3,4 or 5; X is O, N or S; If R ₁And R ₂All be H and X=O, then n can not equal 5.

Those compounds of especially preferred structural formula I representative have contraposition amidine structure, shown in following structural formula Ia:

R wherein ₁, R ₂, R ₃, X, m be identical with the definition among the structural formula I with n.

It itself is new compound that many discoveries can be used for treating or preventing the compound of pneumocystis carinii pneumonia.This class new compound such as following structural formula II define:

Wherein, each R ₁Be H, or two R on same amidino groups ₁The common representative-(CH of base ₂) _m-, m=2 wherein, 3 or 4; R ₂Be H, OCH ₃, NO ₂Or NH ₂R ₃Be H, CH ₃Or CH ₂CH ₃N=2,3,4 or 5; X is O, N or S; Supplementary condition are to work as R ₁And R ₂When all being H, then X represents N or S, and works as R ₂Be H, when X is O, two R then ₁The common representative-(CH of base ₂) _m-, and n=3 or 4.

Especially preferred compound with contraposition amidine structure by the structural formula II representative, shown in minor structure formula IIa:

Wherein, R ₁, R ₂, R ₃, X, m be identical with the definition in the structural formula II with n.In addition, be different from the new compound (but wherein n=6) of structural formula II definition in addition, these compound exhibits go out the effect of anti-PCP, but have higher toxicity.

Giardiasis

Have been found that and to treat giardiasis with five amidines ex hoc genus anne thing.Therefore, the invention provides a kind of method for the treatment of giardiasis, comprise the compound in structural formula I of using effective in cure dosage to ill host, wherein X is O, N or S; R ₁Be H, or two R on the same amidino groups ₁The common representative-(CH of base ₂) _m-, m=2 wherein, 3 or 4; R ₂Be H, NH ₂, OCH ₃, Cl or NO ₂R ₃Be H, CH ₃Or CH ₂CH ₃And n=2-6, or it can make medicinal salt.Preferable is the compound that structural formula II is represented, wherein X, R ₁, R ₂, R ₃, m and n and aforementioned same meaning, or it can make medicinal salt.

Leishmaniasis

Now beat all discovery can be treated leishmaniasis with some five amidine congener.Therefore, the invention provides a kind of method for the treatment of leishmaniasis, comprise the compound in structural formula I of using effective in cure dosage to patient host, wherein X is O, N or S; R ₁Be H, or two R on the same amidino groups ₁Common representative-(CH ₂) _m-, m=2 wherein, 3 or 4; R ₂Be H, NH ₂, OCH ₃, Cl or NO ₂R ₃Be H, CH ₃Or CH ₂CH ₃, n=2-6, supplementary condition are to work as R ₁And R ₂All be H, and during X=O, then n can not equal 5, or it can make medicinal salt.Preferable is the compound that structural formula II is represented, X wherein, R ₁, R ₂, R ₃, m and n and above-mentioned same meaning, or it can make medicinal salt.

Malaria

Have now found that available five amidines ex hoc genus anne thing treatment malaria.Therefore, the invention provides a kind of method for the treatment of malaria, comprise the compound in structural formula I of using effective in cure dosage to ill host, wherein X is O, N or S; R ₁Be H, or two R on same amidino groups ₁Common representative-(CH ₂) _m-, m=2 wherein, 3 or 4; R ₂Be H, NH ₂, OCH ₃, Cl, or NO ₂R ₃Be H, CH ₃, or CH ₂CH ₃, and n=2-6, supplementary condition are to work as R ₁And R ₂All be H, and during X=0, then n can not equal 5; Or it can make medicinal salt.Preferable is compound in the structural formula II, X wherein, R ₁, R ₂, R ₃, m and n such as above-mentioned definition, or it can make medicinal salt.

New compound

In general, the present invention also provides the prescription of medicine, comprising aforementioned structural formula II (or structural formula II new compound a) preferably, or it can make medicinal salt, and physiologically acceptable carrier.The present invention also provides this class new compound or its salt, and they are lyophilized forms, and can be mixed with the medicinal preparation of doing again, as is used for vein or intramuscular injection.

Novel composition

Further, the invention provides a kind of above-mentioned compound or the prescription of its salt and use, as be atomized into particle or droplet and take for sucking to the patient.

New compound

New compound of the present invention is fairly obvious with the difference of prior art compound on constitutional features, and by the structure of this compounds is compared with five amidine structures shown in the structural formula II I, can be easy to determine the difference between them: Wherein Am represents amidino groups.

In one aspect of the invention, owing to replaced two fragrant ether oxygen of examining in the bridge joint group with nitrogen-atoms or sulphur atom, thus new compound and five amidines and its known congener are made a distinction.This class new compound possible constructions formula II (or minor structure formula IIa) represents that wherein X is N or S.In this class example, new compound has the structure that following general formula I Va or IVb represent, or preferablely has a special contraposition amidine structure shown in minor structure formula IVc or the IVd:

Wherein, each R ₁All be H, or two R on same amidino groups ₁Common representative-(CH ₂) _m-wherein m is 2,3 or 4; R ₂Be H, OCH ₃, NH ₂Or NO ₂R ₃Be H, CH ₃Or CH ₂CH ₃N=2,3,4 or 5.As pointing out, there is not such compound in the prior art, be by-N-(CH between two virtue nuclear ₂) _n-N-or-S-(CH ₂) _nThe group of-S-this structure connects, and this exclusive feature handle has the compound of IV structure and those differences in the prior art are come.

Another aspect of the present invention is owing to be connected with a methoxyl group on two virtue nuclears, amino or nitro and new compound of the present invention is distinguished with prior art come.Work as R ₂Be OCH ₃, NH ₂Or NO ₂The time, this compounds by structural formula II representative (or more preferably structural formula II a) and can be by structural formula V, or is preferably represented by the contraposition amidine structure of minor structure formula Va:

Wherein, each R ₁Be H, or two R on same amidino groups ₁Common representative-(CH ₂) _m-, wherein m is 2,3 or 4; R ₂Be OCH ₃, NH ₂Or NO ₂R ₃Be H, CH ₃Or CH ₂CH ₃X is O, N or S; N=2,3,4 or 5.Because methoxyl group, amino or nitro are arranged, therefore be different from five amidines and its congener in this compounds.Find methoxyl group and the amino treatment effectiveness of this compounds aspect anti-Pneumocystis carinii activity that increased.Particularly methoxyl group is greatly enhanced the result of treatment of compound.

In another aspect of this invention, because there is the closed loop derivative of amidino groups in some new compound on two virtue nuclear,, make them different with the compound in the prior art as the tetrahydroglyoxaline ring.This closed loop, for example tetrahydroglyoxaline is by one-(CH ₂) _m-group, for example-CH ₂CH ₂-, the nitrogen-atoms on two amidino groups of bridge joint forms.Reference structure formula II, two R on same amidino groups ₁Common representative-(CH ₂) _m-, m=2 wherein, 3 or 4 Shi Ze have represented this compounds.This compounds is N or S at the X place, and/or works as R ₂Be OCH ₃, NH ₂Or NO ₂The time, still unknown in the prior art.And, when X is 0, and n=2,3 or 4 o'clock these compounds also be unknown in the prior art.But working as X is 0, R ₂Be H, and during n=5, imidazolinium compounds is known.It but is unknown that yet described compound has the treatment effectiveness of anti-Pneumocystis carinii.Have closed loop on the new compound of the present invention, for example during imidazolinyl, be surprisingly found out that the effectiveness of compounds for treating pneumocystis carinii pneumonia significantly strengthens.This class new compound is represented specially by structural formula VI, and minor structure formula VIa then represents more desirable contraposition imidazoline structure: Wherein, R ₂Be H, OCH ₃, NH ₂Or NO ₂R ₃Be H, CH ₃, or CH ₂CH ₃X is O, N or S; N=2,3,4 or 5, if R ₂Be H, then n can not equal 5.The most desirable compound is to work as R ₂=OCH ₃, R ₃=H is when X=0 and n=3, by the compound of minor structure formula VIa representative.Treatment of Pneumocystis carinii and prevention

The aspect of a particularly important of the present invention has provided a kind of method of pneumocystis carinii pneumonia.This and method comprise uses compound in structural formula I with dosage or it can make medicinal salt to the patient who suffers from pneumocystis carinii pneumonia.Before this, five amidines are a kind of in only several its structure compound similar to formula I compound, known it can treat or prevent pneumocystis carinii pneumonia effectively.Only other that know have two amidines that some anti-PCP renders a service be bromo the third two amidines, _ amidine and hydroxyl _ amidine.

Except the method that a kind of pneumocystis carinii pneumonia is provided, the present invention also provide a kind of in the patient that immunity system suffers damage (for example AIDS patient) prevention pneumocystis carinii pneumonia method, this class patient must cross pneumocystis carinii pneumonia at least one time, but does not show symptoms of pneumonia when treatment.Because pneumocystis carinii pneumonia is a kind of disease with special harmfulness for the patient of immune defect, therefore, treat again after sick therewith symptom occurs and compare, had better avoid the outbreak of pneumocystis carinii pneumonia.Therefore, the invention provides a kind of method of preventing pneumocystis carinii pneumonia, comprise the compound of using structural formula I (preferably minor structure formula Ia) to the patient, or it can make medicinal salt with prevention effectiveness dosage.The compound that uses according to this method or the administration form of salt, can be used for the identical of actual therapeutic pneumocystis carinii pneumonia patient.

The present invention another useful aspect, provided a kind of in the patient of the immune defect that never must cross pneumocystis carinii pneumonia the method for prevention initial stage pneumocystis carinii pneumonia.In this respect, a patient who is diagnosed as immune defect, for example AIDS VICTIMS or ARC (AIDS AIDS-related complex AIDS), even before the pneumocystis carinii pneumonia outbreak in the early stage, also can make medicinal salt and avoid or delay infecting with there being prevention to render a service the compound of structural formula I (preferably structural formula Ia) of dosage or its.This compound or salt can be used with the same mode of pneumocystis carinii pneumonia patient and use.The methods of treatment of giardiasis

The present invention also provides the compound of a kind of said structure formula I that has dosage by use or its can make the novel method that medicinal salt is treated giardiasis.Structural formula I comprises five amidines, and its various congeners or derivative, and all these compounds all are aromatics two amidines.

Find that according to the practice of the inventive method at utilization structure formula I, or the best compound of structural formula II, or it be can make medicinal salts for treating giardiasis the time, the curative effect of some compound is better than other compounds.For example, find that five amidines have the medium Lan Shi giardia lamblia that kills and render a service, as the most compounds among the said structure formula I (or II).Especially surprisingly, find that the most virtuous product is a kind of compound with structural formula II definition structure in the scope of the invention, X=O wherein, R ₁And R ₃=H, R ₂=OCH ₃, n=3.During giardiasis, rendeing a service closely similar compound is the compound that structural formula II defines in treatment, X=N wherein, R ₁, R ₂And R ₃=H, n=6.The compound that giardiasis is treated in these compounds and above-mentioned present choosing is compared and can be found, on the curative effect of treatment giardiasis, the compound in the scope of the invention is the same with the product of current use basically.

The methods of treatment of leishmaniasis

The present invention also provides a kind of compound that uses said structure formula I or its can make the method that medicinal salt is treated leishmaniasis.Structural formula I comprises five amidines, and its various congeners or derivative, and all these is aromatics two amidines.

Find that according to the practice of the inventive method at the compound of utilization structure formula I (or preferably structural formula II), or it be can make medicinal salt and treat leishmaniasis the time, the curative effect of some compound is better than other compounds.For example, find that five amidines have medium leishmanicidal effectiveness, as the most compounds among the said structure formula I (or II).Be surprisingly found out that especially the most virtuous product is the compound with structural formula II definition structure in the scope of the invention, X=N wherein, R ₁, R ₂And R ₃=H, n=5 and have the compound of structural formula II definition structure, wherein X=0; R ₁, R ₂And R ₃=H, n=6.These two kinds of compounds are structurally all similar to five amidines.

The method of malaria treatment

The present invention also provides a kind of compound that uses said structure formula I or its can make the novel method that medicinal salt is treated malaria.Structural formula I comprises five amidines, and its various congeners or derivative, and all these is aromatics two amidines.

Find that according to the practice of the inventive method at the compound of utilization structure formula I (or preferably structural formula II), or it be can make medicinal salt and treat malaria the time, the curative effect of some compound is better than other compounds.For example, find that five amidines have medium antiplasmodial effectiveness, as the most compounds among the said structure formula I (or II).Be surprisingly found out that especially within the scope of the present invention, the most virtuous product is No. 106 compound up to now, it has the structure of structural formula II definition, wherein X=N; R ₁, R ₂And R ₃=H; N=5.This compound and five amidines are identical, but the bridge joint Sauerstoffatom of five amidines is replaced by nitrogen-atoms.Especially beat all is that this compound all is compounds effective at chloroquine resistant strain (W2) and mefloquine resistant strain (D6).

Very akin with it compound is the 110th, 113 and No. 116 compound on the plasmodium effectiveness killing.

Dosage and dosage form

Obviously, the dosage that result of treatment is arranged of any specific compound what can be different because of compound, people and different due to illness.With regard to generalized case, dosage will produce the treatment function approximately from about 0.1mg/Kg to 20mg/Kg.But the toxicity problem that high dosage causes can be with dose limitation on lower level, and as the highest 10mg/Kg that is about, this amount is meant the weight of free alkali.The typical doses that is adopted is about about 0.5mg/Kg to 5mg/Kg.The treatment time length is generally once a day, and the symptom that continues up to patient is eliminated.According to the severe degree that individual patients infects, this time length may continue for 2-3 weeks, and is perhaps longer.

According to present method, compound in structural formula I, or its pharmaceutically adoptable salt can adopt oral or suck as solid, maybe can adopt oral or do muscle or intravenous injection by solution, suspension or emulsion.In addition, this compound or salt also can carry out muscle or intravenous injection by liposome suspension.When medicinal suction, compound and salt should be with many solid particulates or droplet forms, and granular size is about 0.5 to 5 μ m, preferably is about 1 to 2 μ m.

Novel composition

The present invention also provides a kind of new medicinal compositions that is used for muscle or used for intravenous injection.Medicinal compositions is made of compound or its pharmacy acceptable salt of structural formula II (preferably minor structure formula IIa), uses any pharmaceutically adoptable carrier.If required is solution, so with respect to water-soluble compound or salt, selection water is carrier; With respect to water-fast compound or salt, select organic carrier for use, for example glycerine, propylene glycol, polyoxyethylene glycol or its mixture may be suitable.Under one situation of back, organic carrier can comprise a large amount of water.Under two kinds of situations, in any suitable manner solution is carried out sterilising treatment then in front, the most handy 0.22 μ m filter filters.After the sterilising treatment, solution is poured in the suitable containers, for example the pyrogen-free phial.Then to the bottle sealing of sterilizing, and if necessary vial content is carried out freeze-drying.

Except the compound or its salt of structural formula II (preferably minor structure formula IIa), medicinal compositions also can comprise other additives, regulates additive as the pH value.Especially, useful pH regulator agent comprises acid or alkali or buffer reagent, as Sodium.alpha.-hydroxypropionate, sodium-acetate or gluconic acid sodium salt.Moreover composition can comprise microbiological antiseptic.The available microbiological antiseptic has methyl p-hydroxybenzoate, propylparaben and benzyl alcohol.When being placed multi-dose vials, said composition generally all uses microbiological antiseptic.Certainly, as above-mentioned, medicinal compositions of the present invention can carry out freeze-drying with well-known prior art.

Another aspect of the present invention is, a kind of injectable, stable sterilization composition that is made of the compound or its salt of structural formula II (being preferably minor structure formula IIa) is provided, and is contained in the sealed vessel with unitary dose.This compound or salt provide with lyophilized form, and this lyophilized products can reconstitute the fluid composition that is applicable to the human injection with acceptable carrier pharmaceutically.The typical unit doses form contains compound or the salt about about 10mg to 10g.When compound or salt are water insoluble basically, can use the physiologically acceptable emulsifying agent of q.s so that in aqueous carrier emulsified compound or salt.A kind of available emulsifying agent is a phosphatidyl choline.

According to the present invention, can prepare other medicinal compositionss, for example aqueous emulsion from the compound or its salt of water-fast structural formula II (preferably minor structure formula IIa).In this case, composition will comprise the pharmaceutically acceptable emulsifying agent of q.s, so that make the compound or the emulsifying salt of the structural formula II (preferably minor structure formula IIa) of aequum.Especially the available emulsifying agent has phosphatidyl choline and Yelkin TTS.

In addition, the invention provides the liposome prescription of the compound or its salt of structural formula II (preferably minor structure formula IIa).The technology that forms lipid suspension is known prior art.When the compound or its salt of structural formula II is a kind of water-soluble salt, use traditional liposome technology, it can be mixed in the lipid carrier.In this case, because that compound or salt have is water-soluble, compound or salt will enter the hydrophilic centre or the nuclear of liposome basically.The fat layer that uses is any traditional composition, and can comprise cholesterol or not have cholesterol.When the compound that relates to or salt are water insoluble, to use traditional liposome again and form technology, salt can enter the hydrophobic class lipid bilayer that forms liposome structure basically.In these two kinds, under arbitrary situation,, can reduce the liposome that is produced dimensionally by adopting the sonication and the homogenate technology of standard.

Certainly, but comprise structural formula II compound or its salt the lipid prescription also freeze-drying produce lyophilized products, this lyophilized products can be used pharmaceutically acceptable carrier, for example water reconstitutes liposome suspension.

In another aspect of this invention, provide a kind of formula of medicine that is suitable for sucking, i.e. aerosol.These prescriptions comprise required compound or the solution of its salt or many solid particulates of suspension or this compound or salt of structural formula II (preferably minor structure formula IIa).Required prescription can be placed the loculus container and with its spraying.Nebulization can produce by pressurized air or by ultrasonic energy, forms the many liquid droplets or the solid particulate that contain compound or salt.The granular size that liquid droplet or solid particulate should have is approximately in the scope about 0.5 μ m to 5 μ m.Solid particulate can pass through to use known suitable method, for example micronization, and the solid chemical compound of processing treatment structural formula II or its salt are produced.Best, the size of solid particulate or little droplet should be about 1-2 μ m.In this respect, Shang Yong atomizer can reach this purpose fully.

Preferably, when the formula of medicine that is suitable for taking as aerosol is liquid form, in the aqueous carrier of bag, prescription will comprise water-soluble compound or its salt of structural formula II (being preferably minor structure formula IIa).Can use tensio-active agent to reduce the surface tension of filling a prescription, so that when atomizing, be enough to make the little droplet of formation in required magnitude range.

As mentioned above, the invention provides water-soluble and water-fast compound and salt.As the term that uses in this specification sheets " water-soluble ", its connotation is defined as any composition, and its water-soluble amount is about 50mg/ml or bigger.In addition, as the term " water-fast " that uses in this manual, its connotation is defined as any composition, and its solubleness in water is lower than 20mg/ml.In some application scenario, need water-soluble compound or salt, and also may need water-fast compound or salt in other application scenarios.

Building-up process

Compound used therefor of the present invention, no matter be known or new compound, all available general known method or method of the prior art are synthetic.Therefore, the synthetic method of these compounds of great majority is not done very detailed explanation at this.Hereinafter related route of synthesis general introduction is in order to help ordinary skill to select known suitable building-up process for use for different classes of compound.

In general, can be used for some reaction sketches of The compounds of this invention synthetic and be shown in figure I-III.As scheme shown in the I, in structural formula I, n=2-5 (being during n=6), R in order to compare ₁=H, X=O and R ₂=H or OH ₃This compound can be produced by carry out alkylation when suitable (replace with methoxyl group) with dibromo alkane 4-hydroxybenzonitrile, the general technology of using people (J.Med.Chem.16:970,1973) such as Geratz is carried out, so that obtain the corresponding cyano group analogue of similar structures formula II compound.Then, the cyano group analogue can be synthetic through the PinnerShi amidine, to produce required product.In addition, imidazoles (imidate) also is to obtain by above-mentioned alkylation, and it can reflux with quadrol and produce tetrahydroglyoxaline product of the present invention, and this product is as two R ₁Base representative-CH together on amidino groups ₂CH ₂In-time, can represent by structural formula I.Moreover, the cyano compound that is obtained by aforementioned alkylated reaction can further reaction obtains the dinitrobenzene di-nitrile compound by carrying out nitrated with acetyl nitrate in trifluoroacetic acid, the PinnerShi amidine of this compound application of aforementioned is synthetic then, can be converted to corresponding amidine.For comparison purpose, can study resulting dinitrobenzene two amidine compounds.These dinitrobenzene two amidine compounds can further be used H ₂Transform with Pd/C, generate useful corresponding diamino two amidines of the present invention.

As scheme as shown in the II, with structural formula I (X=N wherein, R ₂=H) diazo compound derivative of representative can be by the nucleophilic displacement reaction of 4-fluorophenyl cyanogen and diamino alkane, and the PinnerShi amidine that carries out of continuing is synthesized and synthesizes.Equally, 4-fluorine-3-cyanide-nitrophenyl can react with the diamino paraffinic hydrocarbons, thereby generates corresponding cyano derivative.Then, relative solubility is depended in this intermediate product to the conversion of the finished product.Referring to figure III, the figure illustrates compound (X=N wherein, R in this respect ₂=NH ₂Synthesizing and n=2), promptly under 25 ℃ of conditions, make the reacting ethylenediamine of 4-chlorine-3-cyanide-nitrophenyl with ten times of surpluses, generate corresponding single derivative, then, this derivative can be formed corresponding amine by catalytic reduction, that continues carries out second time nucleophilic displacement reaction with 4-chlorine-3-cyanide-nitrophenyl and forms corresponding dicyano mono amino mononitrile, then, can make it carry out the synthetic and final reduction of PinnerShi amidine, generate needed diamino two amidine products.

Equally, compound in structural formula I has also been represented a synthetic result, wherein, and X=N, R ₂=NH ₂And n=4 or 6, in this was synthetic, 4-chlorine-3-cyanide-nitrophenyl produced a kind of intermediate product that is insoluble to any suitable solvent with the intermediate product cyano derivative that corresponding diamino paraffinic hydrocarbons reaction obtains, so that change into corresponding two amidine derivatives.In this case, nitro can be reduced into amino, changes into two required arteries and veins then, and is described as approach 2 among the figure II.The compound that other are same, but n=3 or 5 wherein are slightly soluble in the dioxan, therefore can directly be transformed into two amidines, produce final needed diamino two amidine compounds before nitroreduction.

As mentioned above, the used compound of the present invention can be used as pharmacy acceptable salt provides.Such salt comprises gluconate, lactic acid salt, acetate, tartrate, Citrate trianion, phosphoric acid salt, borate, nitrate, vitriol and hydrochloric acid salt.

In general, salt of the present invention can prepare with surpassing 2 normal required acid-responss slightly by the amidine alkali cpd in solution.After reaction is finished, add the undissolved therein solvent of an amount of salt, salt is crystallized out from solution.

By limiting examples hereinafter the present invention is further set forth.

Example 1-44 (comprising comparative example)

According to the synthetic compound of suitable procedure discussed above with structural formula I structure.In figure I-III, provide the reaction sketch, shown the method that is used for synthetic each example compound one by one.The synthetic compound is shown in Table I A, and the ultimate analysis of compound and fusing point are shown in Table I B.The experimental arrangement of use standard carries out toxicity test to most compounds in rat, toxicity test the results are shown in Table X II.Example 31,32 and 34 compound can be synthetic with aforesaid general method, but not synthetic these compounds.

Compound is to the test of Pneumocystis carinii result of treatment

In Sprague-Dawley rat to introducing and the treatment of Pneumocystis carinii

Male Sprague-Dawley rat of feeding, definite virus-free, weight 150-200 grams derive from the Hilltop laboratory, and dress are opened respectively.Animal once arrive set about feeding low (8%) albumen food (ICN Biochemicals, Cincinnati, OH) and contain the drinking-water of tsiklomitsin (0.5mg/ml) and dexamethasone (1.0g/ml).Thereafter 8 weeks were carried out such treatment, and the liquid of monitoring picked-up every day and claim the weight of animal weekly.When animal absorbs too many liquid, drinking-water is diluted, poison to avoid cortisone.Since the 6th week, animal is divided into one group of per 8 animal, and except as otherwise noted, afterwards 14 days, every day with the dosage of 10mg/Kg by the intravenous injection test compound.

At the 8th weekend, suck the execution animal by trichloromethane, for tissue abrasion, sterilization ground takes out left lung, places the aseptic HanKShi balanced salt solution (HBSS) of no calcium or magnesium.Make right lung expansion and taking-up carry out histology GMS chromoscopy in the original place with 10% prescription.

The organized processing program

The induced lung that can not handle is at once carried out quick-frozen and is stored under-70 ℃ of temperature.When preparing to handle, from refrigerator, take out tissue, and make its quick-thawing in 25 ℃ of water-baths.Then, lung is cut into small pieces, grinds by the #60 silk screen with the glass pestle.The lung of breaking into pieces is suspended in 30 seconds of 10ml HBSS solution mesoscale eddies.With suspension centrifugal 10 minutes with 60 * g, abandon precipitation and supernatant is moved in another pipe, with 150 * g centrifugal 10 minutes, to remove remaining cell debris.Then, again with the centrifugal supernatant that goes out centrifugal 10 minutes, so that make lung sac worm precipitation with 10,000 * g.Precipitation is resuspended among the HBSS of 2ml and dyes.On clean slide, prepare slide glass by placing 10 μ l lung suspension, make it air-dry.(Kodak Chemicals, Rochester is NY) with slide glass dyeing, so that show the scrotiform shape that this is biological to use cresyl typical case purple then.

Statistical research

For each lung suspension, 20 powerful microscope visuals field are totalized, and calculate the average number of capsule.Test-results

Table II

Can be used for treating the compound of PCP in the example 4 of the present invention, and common contraposition shape and position shape five amidine compare test.The compound of example 4 is different from five amidines (contraposition shape), and what wherein connect two virtue nuclears is-(CH ₂) ₃-base rather than five amidines-(CH ₂) ₅-base.The compound of finding example 4 is being better than a contraposition or position shape five amidine aspect the degree of control PCP infection.

Table III

The two kinds of new compounds and five amidines that the present invention are had structural formula II (and minor structure formula IIa) compare test, are different from five amidines on this compound structure, and the one, because R ₂Be methoxyl group, and another is because R ₂Be an amino (being respectively example 15 and 14).Also comprised a kind of known compound under study for action, its structure is different from five amidines, and wherein n is 6, represents six amidines (example 20).The discovery that the people is taken aback is that all the 3 kinds differences with five amidine structures have all been improved the effect of pneumocystis carinii pneumonia.

Table IV

The fourth amidine (example 9) of structural formula I (and minor structure formula Ia) is compared with five amidines and two analogue, first in the scope of structural formula II (minor structure formula IIa), R wherein ₂Represent NH ₂(example 19), second structural formula I (minor structure formula Ia) and II (in minor structure formula II scope a), R wherein ₂=NO ₂Under the situation of amino-substituted compounds (example 19), the ether oxygen of five amidines (position X) is also replaced by nitrogen.The discovery that is filled with amazement is to be better than five amidines greatly fourth amidine aspect the control pneumocystis carinii pneumonia.Other two compound comparisons are shone, but are being not so good as five amidines aspect the control pneumocystis carinii pneumonia.

Table V

Structural formula II (and minor structure formula IIa, example 7,8,17 and 18) four compounds are similar to five amidines of n=3-5, but substituted O (position X) with N, the derivative that is n=6 with they and five amidines and middle abutment chain length thereof compares test, has produced higher toxicity.New compound (n=5 with nitro substituent, example 18) be better than contrast, but be not so good as five amidines on the effect, and the new compound of example 7,8 and 17 is better than or equal five amidines, and the compound of n=3 (example 7) is the most effective, and the compound of n=5 (example 17) effect is the poorest.The derivative of n=6 can be compared with five amidines, but toxic influence (seeing Table XIII).

Table VI

Four kinds of new compounds of structural formula II (minor structure formula IIa, example 3,5,10 and 14) are aspect pneumocystis carinii pneumonia effect, compare test with five amidines, these compounds all have amino substituting group on virtue nuclear, as shown in the structural formula II (minor structure formula IIa), and R wherein ₂=NH ₂Moreover in all cases, the linking group of virtue nuclear comprises two ether oxygenses, as representative when the X=O.The chain length of bridged bond alkyl changes in 2-5, for purpose relatively, has used another compound identical with compound in the structural formula II, and wherein alkyl chain length is 6 (as representative when the n=6).Compound in structural formula II (minor structure formula IIa) scope all is being better than contrast greatly aspect the pneumocystis carinii pneumonia, and big activation is compared with five amidines on effect.The compound of n=6 obviously is worse than other compounds or five amidines and has toxicity in the example 22 on pneumocystis carinii pneumonia effect, sees Table XIII.

Table VII

Be purpose relatively, aspect the pneumocystis carinii pneumonia effect to five amidines and four kinds wherein amidino groups be between the analogue of position analyze.Between the carbon chain lengths of linking group of position amidino groups analogue in 3-6, change, as representative when the n=3-6.In these position five amidines, though none has the result of treatment the contraposition of resembling five amidines, they are better than contrast.What is particularly worth mentioning is that, have meta-compound than the long abutment of short chain (n=3,4) be better than between position five amidines.For purpose relatively, use the blocking-up amidine the same to test equally, find aspect result of treatment, quite or slightly to be better than five amidines and to handle five amidines with five amidines with five amidines.

Table VIII

With containing the compound of methoxyl group in the structural formula II (minor structure formula IIa, example 6,11 and 15),, comparing with five amidines aspect the result of treatment of pneumocystis carinii pneumonia with 5mg/Kg.Said compound is with R ₂=OCH ₃The time structural formula II (minor structure formula II a) is represented.Moreover these specialized compounds comprise oxygen in the bridge joint group of two virtue nuclears, shown in the structural formula II of X=O.The variation length of abutment carbochain is 3-5, shown in n=3-5 o'clock.Also comprised two kinds of compounds under study for action, wherein virtue nuclear is gone up and is replaced by the chlorine atom, as R ₂Shown in the structural formula II a of=Cl, it is better than contrast slightly in the pneumocystis carinii pneumonia when 2.5mg/Kg dosage, but more very different than five amidines.The compound that comprises methoxyl group when 1/2nd dosage significantly better than contrast, and, according to the length of chain, be worse than, be equivalent to aspect the pneumocystis carinii pneumonia or significantly better than five amidines with 1/2nd dosage of five amidines.To the compound that this methoxyl group replaces, the short more effect of chain length is good more, along with chain length becomes the large effect variation.

Table I X

Four kinds of compounds in the structural formula II scope (example 33,41,42 and 44) are being compared with five amidines aspect the effect of pneumocystis carinii pneumonia.In a kind of situation (example 33), new compound of the present invention is at virtue nuclear (R ₂=NH ₂) on comprise an amino substituting group and in the groups that are connected two virtue nuclears, nitrogen-atoms (X=N) arranged and to have short abutment alkyl chain length (n=2).This compound comparison is shone, but poorer than five amidines.Also the dosage with 2.5mg/Kg has compared three kinds of compounds (example 41,42 and 44), and wherein amidine nitrogen base is by etheno connection formation tetrahydroglyoxaline.The compound of example 44 has methoxy substituting group (R on virtue nuclear ₂=OCH ₃), and ought virtue nuclear connection base be-O (CH ₂) ₃During O-(is representative with X=0 and n=3), find that its pneumocystis carinii is very effective, all much better when having only 1/4th dosage than five amidines.Two similar compounds are also tested, and they have methyl substituents on imidazolinyl.Find that such methyl existence has obviously weakened compound functions, but still be better than contrast.Compound 44 is represented most preferred embodiment of the present invention, and wherein under five amidines, 1/4th dosage conditions, the effect of pneumocystis carinii pneumonia is very good.

Table X

Just to the treatment function of pneumocystis carinii pneumonia, to 6 kinds in structural formula II a scope the new compound of (example 35,36,37,38,40 and 43) test.Two new compounds (example 36 and 37) have the amino (R of replacement on virtue nuclear ₂=NH ₂), and in virtue nuclear abutment, have nitrogen-atoms (X=N).These compounds are compared with similar new compound, and wherein the substituting group on virtue nuclear is nitro (example 35 is tested with 5mg/Kg, and example 38 is tested with 2.5mg/Kg), and the chain length of abutment changes (n=3-4) in 3-4.Find that amino substitution compound is better than the compound that nitro replaces, although amino-substituted compounds is tested under higher dose levels.In addition, (two R have wherein been tested and have had imidazolinyl ₁Base is-CH ₂CH ₂-) structural formula II a in two compounds (example 40 and 43), a kind of compound (example 43) has the methoxyl group (R on virtue nuclear ₂=OCH ₃), and the group of bridge joint virtue nuclear is-O (CH ₂) ₅O-, in another kind of situation (example 40), on virtue nuclear, there is not replacement, abutment is-O (CH ₂) ₄O-.Find that two such compounds all are better than contrast significantly, even the compound of example 43 is to test under the lower level of 5mg/Kg.

Table X I

For with fourth amidine (R ₁=H, R ₂=H, X=0, function n=5) and five amidines compare, and are 10mg/Kg at dosage, under the condition of 1mg/Kg and 0.1mg/Kg, just for the effect of pneumocystis carinii pneumonia fourth amidine and five amidines are carried out the logarithm comparison test.Under the condition of 10mg/Kg dosage, the fourth amidine is better than five amidines greatly, has confirmed the result described in the Table IV of front.But, when dosage level drops to 1mg/Kg or 0.1mg/Kg, basic indistinction between two kinds of compounds.

Brief summary

The synthesis result of foregoing in vivo test is listed among the Table X II of back.Although because the contrast difference between the test can cause synthesis result is not very definite, its a kind of useful instrument of can yet be regarded as has also fully been summed up effectiveness of the present invention.

Example 45-80

Test at the Lan Shi giardia lamblia

According to non-limitative example hereinafter the present invention is further set forth.Example 45-88 (comprising comparative example).

The compound that use belongs in structural formula I (and II) scope is tested, and they have at Table X IV to the structure shown in the Table X VI.For these compounds are tested at the Lan Shi giardia lamblia, carry out following general procedure.The sterile culture thing of Lan Shi giardia lamblia trophont.

With Lan Shi giardia lamblia WB strain (ATCC ^#30957) grow in TY-S-33 medium (people such as Diamond of filtration sterilization, 1978) in, this medium has carried out modifying (Keister by adding bile, 1983), and the calf serum, 50 μ g/ml penbritins (Sigma) and the 50 μ g/ml vitriol gentamicins (Sigma) that contain 10% thermal inactivation.The original seed culture of trophont is grown under 37 ℃ of conditions, in 13 * 100mm nut borosilicate glass test tube.Every 72 hours,, culture tube organism was cultivated again in 5 minutes by being cooled off in ice-water bath.Toppling over cooling tube with strength takes out trophont from glass test tube.Organism number during placement 10 μ l suspension are determined every milliliter on hematimeter.Logarithmic phase culture with about 5 * 104 moves in the new medium.

Chemotherapeutic

Use process described in detail in the preamble, used five amidines and five amidine analogues in synthetic this research.Obtain Metronidazole plain BP.98 99, quinacrine hydrochloride and Trichofuron from Sigma company.The protospecies solution of preparation 2mM in TYI-S-33 medium of modifying, and further with the susceptibility of medium with the mensuration medicine.

Little culture of Lan Shi giardia lamblia

From the original seed culture of logarithmic phase, obtain Lan Shi giardia lamblia trophont.To contain TYI-S-33 medium that 100 μ l of 2.5 * 104 trophonts modify inserts in the hole of the tissue culturing plate (Costar) at the bottom of the 96 hole U types.Tissue culturing plate is placed in the plastics casing, and it is in the anaerobic environment, regulates this plastics casing and makes it make nitrogen pass through this box by import and outlet conduit, and this pipeline can clamp and close.Nitrogen by box 2 minutes, and is placed container under 37 ℃ of conditions and to pass to 5%CO ₂The incubator of air in.Viability ³H-thymidine mixes mensuration

With different concns ³H-thymidine (0.5,1.0,1.5 and 2.0 μ Ci/ hole) added at interval with 2 hours and contains in the hole of 2.5 * 104 Lan Shi giardia lamblia trophonts, so that make it contact 2-24 hours with trophont.After insulation 24 hours, by washing with strength, with smashing the cell harvestor collecting cell to pieces with ice-cold HanKShi balanced salt solution more.With sample collection on Whatman glass microfiber paper.The paper dry air places 7ml flicker bottle with sample, and determines by liquid flashing counting ³Mixing of H-thymidine.5 double counting mean values by each time point and concentration.With these data determine Best Times and ³The concentration of H-thymidine is for use in measuring drug susceptibility.

Determination of drug sensitivity

TYI-S-33 medium that 100 μ l are contained the modification of 2.5 * 104 trophonts places each hole of 96 hole U base plates.This plate is incubated 24 hours down in anaerobic condition.After 24 hours, from the 2mM protospecies solution, prepare test compound, and every kind of solution is added 100 μ l, in the hole, to form the concentration of 1,10,200 and 1000 μ M.Adding medicine after 6 hours, in the TYI-S-33 that modifies, adding 10 μ l150 μ Ci/ml's ³H-thymidine (SA=10Ci/ micromole) solution is to obtain ultimate density 1.5 μ Ci/ holes.Collecting cell after being incubated 18 hours again.Each drug level is done 5 repetitions, and definite mean value.Do not determine with adding biological control wells ³The non-specific combination of H-thymidine and primitive fiber paper.By relatively being added with the compound hole and not adding in the medicine control wells ³H-thymidine mix activity with the confirmed test compound.Determine to suppress 50% ³H-thymidine (IC ₅₀) mix required drug level (Chou, 1974).

Test-results

With the results are shown among Table X IV-XVI of the effect of compound in preceding method mensuration said structure formula I or the II scope in opposing Lan Shi giardia lamblia.From these tables, obviously as seen, work as R at least ₂=NH ₂The time, very short chain length (n=2) effect of abutment is the poorest.But, when n=3, obtained very effective result, no matter R ²=H, NH ₂Or OCH ₃And work as R ₂=OCH ₃, and R ₁And R ₃, obtain best result at=1 o'clock.Really meaningfully, find according to test of the present invention, in treatment Lan Shi giardia lamblia the second optimizing compound by structural formula I definition, X=N wherein, R ₁, R ₂And R ₃=H, and n=6 have showed the group that two virtues of a quite long bridge joint are examined.

By Table X IV as seen, a position amidine can be compared with the contraposition amidine, though when the length of bridge joint virtue nuclear group during in n=4-6 scope, active what are poorer.But when n=3, a position amidine far is worse than its corresponding contraposition amidine.

By Table X V as seen, the compound of structural formula II, the compound that promptly amidino groups is transformed into tetrahydroglyoxaline is generally speaking a little than its single amidine homologue weak effect slightly.Therefore, example 5 compares with example 31 and shows that the tetrahydroglyoxaline of example 31 is poorer slightly than the amidine counterpart effect of example 5.Table X V clearlys show that the substituting group on the imidazolinyl (for example using methyl) can cause effect sharply to descend.Table X VII has comprised the efficacy data of present selected 3 kinds of compounds to treatment Lan Shi giardia lamblia.The effect of these compounds is compared with optimizing compound efficacy data of the present invention, and to have any different on the effect also be extremely small even can see.

Example 81-98 is at leishmanial test

According to limiting examples hereinafter the present invention is further set forth.

Example 81-89

Use belongs to the compound in structural formula I (and II) scope, and they have at Table X VII to the structure shown in the Table X X, test at leishmania for making these compounds, carry out following general program.

Chemotherapeutic

Use synthetic five amidines of above-mentioned detailed process and be used for five amidine analogues of this test.

The active vitro Screening of anti-leishmania

Protomonad with leishmania mexicana (Leishmania mexicanaamazonensis) strain MHOM/BR/73/M2269 (WR669), in the SchneiderShi fruit bat medium of calf serum that is added with 20% thermal inactivation (GIBCO) and 100 μ g/ml vitriol gentamicins, be cultured to initial stage logarithmic phase (parasitology 76:309,1978).Measure medium and add 10% calf serum for the Schneider medium.The compound of the present invention that is used for of serial dilution is suspended in the mensuration medium, and preparation there are 96 hole microtiter plates of repeated rows.(200 μ l) adds in each hole with 2.5 * 106 cells/ml with parasite suspension, and with each dull and stereotyped sealing, is incubated under 25 ℃ of air conditionses.After 24 hours, the adding methyl- ³It is 1-2 μ Ci/ holes that H thymidine (20Ci/ micromole) makes it.After 18 hours, with the Skatron cell harvestor with cell harvesting to the glass microfiber filter.The flushing filter is also dry, with Beckman LS3801 scintillation counter counting.By non-linear regression method, will mix ³The data of H-thymidine are created as logarithm concentration-response function, and determine to suppress 50% ³H-thymidine mixes required drug level (AntimicrobAgents Chemother.16:710,1979; Exper.parasitol.66:86,1988).The results are shown among the Table X VIII-XX of back.

Test-results

With the results are shown among Table X VIII-XX of the effect of compound in the treatment leishmania in aforesaid method mensuration front structure formula I (or II) scope, these tables have showed that contraposition amidine, a position amidine and contraposition tetrahydroglyoxaline all have anti-leishmanial activity.Compound 88 and compound 89 have shown the anti-leishmania activity of height.Compound 88 is except having replaced with nitrogen-atoms the bridge joint Sauerstoffatom, and is identical with five amidines.Compound 89 (six amidines) except comprising a methylene abutment more, and is identical with five amidines.

IX finds out from Table X, and a position amidine can be compared with the contraposition amidine, although they have to a certain degree reduction demonstrating activity in relatively with the contraposition amidine.

Can see from Table X X, the compound of structural formula II, the compound that promptly amidino groups is transformed into tetrahydroglyoxaline generally speaking is equivalent to their amidine homologue basically.Example 99-116

The plasmodium activity

According to limiting examples hereinafter the present invention is further set forth.

Use belongs to the compound in structural formula I (and II) scope, and they have in the structure shown in the Table X XI to XXIII.For testing the effect of these compounds in the treatment malaria, under isolated condition, carry out following general testing sequence.

Chemotherapeutic

Use synthetic five amidines of above-mentioned detailed process and be used for five amidine analogues of this test.

Determination of drug sensitivity

Plasmodium falciparum strain W2 (chloroquine resistance, mefloquine susceptibility) and D6 (mefloquine resistance, chloroquine susceptibility) are being contained RPMI-1640,25mM HEPES, 25mM NaHCO ₃Freeze in the human plasma with 10% (V/V) aquatic foods and to cultivate.The test compound that contains serial dilution, contain the hole that 0.2% to 0.4% parasitic erythrocytic 1.5% red blood corpuscle suspension places 96 hole microtiter plates, every hole 200 μ l.Then flat board is put into the anaerobism container, with nitrogen wash and 37 ℃ of insulations.After 24 hours, add with every hole 25 μ l ³H-xanthoglobulin (1mCi/ml), and as above-mentionedly make dull and stereotyped insulation.After 42 hours, with smashing the cell harvestor collecting cell to pieces on glass fiber paper more.The use scintillation counter is determined ³H one hypoxanthic mixing.By relatively under different pharmaceutical concentration ³H-hypoxanthic mixing, activity (Antimicrob.Agents Chemother.16:710,2979 of the non-linear regression analysis confirmed test compound of using a computer; Am.J.Trop.Med.Hygiene 34:209,1985).The results are shown among Table X IX-XXI.

Test-results

Measure the results are shown among Table X XI-XXIII of the effect of compound in the treatment malaria in structural formula I (or II) scope above with aforesaid method, these tables have showed that contraposition amidine, a position amidine and contraposition tetrahydroglyoxaline all have antimalarial protozoon activity.According to practice of the present invention, the optimizing compound that is used for the treatment of two kinds of plasmodium falciparum strains is No. 106 compounds, and it is defined by structural formula I, X=N wherein, R ₁, R ₂And R ₃=H, and n=5.

Table X XII shows that a position amidine can be compared with the contraposition amidine, although from mean value, they are more or less too late contraposition amidine on activity.

Can see from Table X XIII, the compound of structural formula II, the compound that promptly amidino groups is converted to tetrahydroglyoxaline generally speaking is equivalent to their amidine homologue basically.

Table I A

# X n R ₂1. O 2 H2. O 2 NO ₂3. O 2 NH ₂4. O 3 H5. O 3 NH ₂6. O 3 OCH ₃7. N 3 H8. N 4 H9. O 4 H10. O 4 NH ₂

Table I A (continuing)

# X n R ₂11. O 4 OCH ₃12. O 5 H13. O 5 NO ₂14. O 5 NH ₂15. O 5 OCH ₃16. O 5 Br17. N 5 H18. N 5 NO ₂19. N 5 NH ₂20. O 6 H21. O 6 NO ₂22. O 6 NH ₂23. N 6 NH ₂24. O 4 Cl25. O 5 Cl

Table I A (continuing)

# X n26. O 327. O 428. O 529. O 6

Table I A (continuing)

# X n R ₂31. N 2 H32. N 2 NO ₂33. N 2 NH ₂34. O 3 NO ₂35. N 3 NO ₂36. N 3 NH ₂37. N 4 NH ₂38. O 4 NO ₂39. N 6 H

Table I A (continuing)

# X n R ₂ R ₃30. O 5 H H40. O 4 H H41. O 4 H CH ₃42. O 5 H CH ₃43. O 5 OCH ₃ H44. O 3 OCH ₃ H

Table I B

The ultimate analysis of example 1-44 and the ultimate analysis of fusing point compound

The M.P. productive rate number analysis structural formula that records that calculates

C H N C H N1 C ₁₆H ₁₈N ₄O ₂/2HCl 49.14 5.72 14.33 49.18 5.75 14.28 ＞300 6％2 C ₁₆H ₁₆N ₆O ₆/1.1HCl/1.3H ₂O 42.53 4.39 18.60 42.83 4.01 17.07 287 39％3 C ₁₆H ₂₀N ₆O ₂/4HCl/0.4H ₂O 39.92 5.19 17.46 40.21 5.46 17.08 ＞300 26％4 C ₁₇H ₂₀N ₄O ₂/2HCl/1.3H ₂O 49.96 6.07 13.71 49.96 6.10 13.59 Dec?at?144-145 82％5 C ₁₇H ₂₂N ₆O ₂/4HCl/0.6E1OH 40.81 5.98 15.69 40.72 5.62 15.31 Dec?at?273 6％6 C ₁₉H ₂₄N ₄O ₄/2HCl 51.24 5.88 12.58 51.04 5.91 12.50 293 76％7 C ₁₇H ₂₂N ₆/2HCl 53.27 6.31 21.92 53.11 6.39 21.96 298-300 22％8 C ₁₈H ₂₄N ₆/2HCl/0.5H ₂O 53.20 6.70 20.68 53.04 6.73 20.54 ＞300 43％9 C ₁₈H ₂₂N ₄O ₂/2HCl/0.4H ₂O 53.18 6.15 13.78 53.41 6.21 13.41 ＞300 95％10 C ₁₈H ₂₄N ₆O ₂/4HCl/1H ₂O 41.55 5.81 16.15 41.64 5.84 16.08 285 18％11 C ₂₀H ₂₆N ₄O ₄/2HCl/1.2H ₂O 49.94 6.37 11.65 49.98 6.29 11.61 297-299 70％12 C ₁₉H ₂₄N ₄O ₂/2HCl/1H ₂O 52.90 6.51 12.99 52.66 6.29 12.92 247 86％13 C ₁₉H ₂₂N ₆O ₆/2HCl 45.34 4.81 16.70 45.22 4.86 16.64 255 74％14 C ₁₉H ₂₆N ₆O ₂/4HCl/1.6H ₂O 41.86 6.14 15.42 41.83 6.12 15.37 270 40％15 C ₂₁H ₂₈N ₄O ₄/2HCl/2H ₂O 49.51 6.73 11.00 49.57 6.74 10.99 258-259 63％

Table I B (continuing)

The ultimate analysis of example 1-44 and fusing point compound

The calculating ultimate analysis records

M.P. productive rate number analysis structural formula

C H N C H N16 C ₁₉H ₂₂N ₄O ₂Br ₂/2HCl/1H ₂O 38.73 4.45 9.51 38.84 4.64 9.46 254-255 58％17 C ₁₉H ₂₆N ₆2HCl/1.33H ₂O 52.42 7.10 19.30 52.32 7.08 19.17 295 48％18 C ₁₉H ₂₄N ₈O ₄/2HCl/2H ₂O 42.71 5.76 20.12 43.08 5.41 20.07 293-294 39％19 C ₁₉H ₂₈N ₈/4HCl/2H ₂O 41.47 6.59 20.36 41.60 6.55 20.42 300-303 16％20 C ₂₀H ₂₆N ₄O ₂/2HCl/2.5H ₂O 50.91 7.32 11.31 50.94 7.05 11.31 252 69％21 C ₂₀H ₂₄N ₆O ₆/2HCl 46.43 5.07 16.24 46.51 5.05 16.14 Dec?at?287-288?35％22 C ₂₀H ₂₈N ₆O ₂/4HCl/0.6H ₂O 45.49 6.45 15.01 45.13 6.84 14.61 Dec?at?290 18％23 C ₂₀H ₃₀N ₈/4HCl/1.7H ₂O 42.97 6.74 20.05 43.05 6.76 19.97 Dec?at?285 18％24 C ₁₈H ₁₈N ₄O ₂/4HCl 46.18 4.74 11.97 46.17 4.79 11.90 ＞300 43％25 C ₁₉H ₂₀N ₄O ₂/4HCl/1.1H ₂O 45.46 5.26 11.16 45.49 5.23 10.98 247-248 55％26 C ₁₇H ₂₀N ₄O ₂/2HCl/1.9H ₂O 48.67 6.20 13.35 48.64 6.24 13.31 300 42％27 C ₁₈H ₂₂N ₂O ₂/2HCl/1.8H ₂O 50.07 6.44 12.98 50.10 6.46 12.97 257 68％28 C ₁₉H ₂₄N ₄O ₂/2HCl 55.21 6.34 13.55 55.47 6.51 13.15 133-134 51％29 C ₂₀H ₂₆N ₄O ₂/2HCl/0.3H ₂O 55.51 6.66 12.95 55.45 6.67 12.88 268 64％30 C ₂₃H ₂₈H ₄O ₂/2.2H ₂O 54.70 6.87 11.09 54.69 6.87 11.06 147 81％

Table I B (continuing)

The ultimate analysis of example 1-44 and fusing point compound

The M.P. productive rate number analysis structural formula C H N C H N31 Not Syntheslzed32 Not Syntheslzed33 C that the ultimate analysis of calculating records ₁₆H ₂₂N ₈/ 4HCl/1.6H ₂O/0.4EtOH 38.84 6.13 21.57 38.83 6.00 21.81＞300 27%34 Not Synthesized35 C ₁₇H ₂₀N ₈O ₄/ 2HCl/H ₂O 41.56 4.92 22.81 41.71 4.89 22.73 Dec at 295 58%36 C ₁₇H ₂₄N ₈/ 4HCl/1.1H ₂O/0.3EtOH 40.66 6.20 21.55 40.29 6.29 21.21 229 10%37 C ₁₈H ₂₆N ₈/ 4HCl 43.21 6.04 22.40 43.30 6.09 22.36 Deo at 274-275 27%38 C ₁₈H ₂₀N ₆O ₆/ 2HCl 44.18 4.53 17.18 44.08 4.55 17.10 298 26%39 C ₂₀H ₂₈N ₆/ 2HCl/0.3H ₂O 55.76 7.16 19.15 55.80 7.17 19.44 300 54%40 C ₂₂H ₂₆N ₄O ₂/ 2HCl/2.2H ₂O 53.81 6.65 11.4 53.80 6.70 11.36 249-250 76%41 C ₂₄H ₃₀N ₄O ₂/ 1HCl/1.5H ₂O 61.33 7.29 11.92 61.17 7.40 11.75 203-205 15%42 C ₂₅H ₃₂N ₄O ₂/ 2HCl/1H ₂O 58.71 7.09 10.95 58.73 7.16 10.88 203-204 19%43 C ₂₅H ₃₂N ₄O ₄/ 2HCl/2.5H ₂O 52.63 6.89 9.82 52.47 6.85 9.97 175 70%144 C ₂₃H ₂₈N ₄O ₄/ 1.75HCl/2.00H ₂O 52.69 6.49 10.69 52.59 6.38 10.46 252 62%

Table II

Disease degree by histological grade

Number of animals example number 0.5 1234

Contrast 0121 412 C R ₂=H, X=O, n=5,15200

Contraposition amidine 4 I R ₂=H, X=O, n=3,34100

Contraposition amidine 28 C R ₂=H, X=O, n=5,32201

Between the position amidine

Table II (continuing)

Histological grade:

0.5=find capsule＜10 in per two cross sections

1=disperses capsule, 5% of＜lung

2=disperses capsule, 5-10% or little infection focus of lung

3=disperses capsule, and 10-50% lung contains more serious infected zone

The lung of 4=＞50% contains the serious zone of many focal infections

C=compares, and I represents the known compound of a structural formula I, and II represents the compounds of a structural formula II

Table III

Disease degree by histological grade

Number of animals example number 0.5 1234

Contrast 0013 412 C R ₂=H, X=O, n=5 0341 020 C R ₂=H, X=O, n=6 3310 015 II R ₂=OCH ₃, X=O, n=5 2130 014 II R ₂=NH ₂, X=O, n=5 24110

Table III (continuing)

Histological grade:

0.5=find capsule＜10 in per two cross sections

1=disperses capsule, 5% of＜lung

2=disperses to assist 5-10% or little infection focus of lung

3=disperses capsule, and 10-50% lung contains more serious infected zone

The lung of 4=＞50% contains the serious zone of many focal infections

C=compares, and I represents the known compound of a structural formula I, and II represents the compounds of a structural formula II

Table IV

Disease degree by histological grade

Number of animals example number 0.5 1234

Contrast 0024 212 C R ₂=H, X=O, n=5 4400 09 I R ₂=H, X=O, n=4 8000 019 II R ₂=NH ₂, X=N, n=5 0242 013 II R ₂=NO ₂, X=O, n=5 32300

Table IV (continuing)

Histological grade:

0.5=find capsule＜10 in per two cross sections

1=disperses capsule, 5% of＜lung

2=disperses capsule, 5-10% or little infection focus of lung

3=disperses capsule, and 10-50% lung contains more serious infected zone

The lung of 4=＞50% contains the serious zone of many focal infections

C=compares, and I represents the known compound of a structural formula I, and II represents the compounds of a structural formula II

Table V

Disease degree by histological grade

Number of animals example number 0.5 1234

Contrast 1131 212 C R ₂=H, X=O, n=5 2131 07 II R ₂=H, X=N, n=3 5120 08 II R ₂=H, X=N, n=4 3100 017 II R ₂=H, X=N, n=5 1231 039 C R ₂=H, X=N, n=6 2100 021 C R ₂=NO ₂, X=O, n=6 4110 018 II R ₂=NO ₂, X=N, n=5 1302 123 C R ₂=NH ₂, X=N, n=6 31201

Table V (continuing)

Histological grade:

0.5=find capsule＜10 in per two cross sections

1=disperses capsule, 5% of＜lung

2=disperses capsule, 5-10% or little infection focus of lung

3=disperses capsule, and 10-50% lung contains more serious infected zone

The lung of 4=＞50 contains the serious zone of many focal infections

C=compares, and I represents the known compound of a structural formula I, and II represents the compounds of a structural formula II

Table VI

Disease degree by histological grade

Number of animals example number 0.5 1234

Contrast 0004 412 C R ₂=H, X=O, n=5 5300 03 II R ₂=NH ₂, X=O, n=2 4400 05 II R ₂=NH ₂, X=O, n=3 2410 010 II R ₂=NH ₂, X=O, n=4 2510 014 II R ₂=NH ₂, X=O, n=5 6100 022 C R ₂=NH ₂, X=O, n=6 01430

Table VI (continuing)

Histological grade:

0.5=find capsule＜10 in per two cross sections

1=disperses capsule, 5% of＜lung

2=disperses capsule, 5-10% or little infection focus of lung

3=disperses capsule, and 10-50% lung contains more serious infected zone

The lung of 4=＞50% contains the serious zone of many focal infections

C=compares, and I represents the known compound of a structural formula I, and II represents the compounds of a structural formula II

Table VII

Disease degree by histological grade

Number of animals example number 0.5 1234

Contrast 0013 412 C, five amidines, 4220 026 I n=3,2141 027 I n=4 between Am=, 2113 028 C n=5 between Am=, 1142 029 C n=6 between An=, between Am=14120---I n=5,53000 of Am=blocking-up

Table VII (continuing)

Histological grade:

0.5=find capsule＜10 in per two cross sections

1=disperses capsule, 5% of＜lung

2=disperses to assist 5-10% or little infection focus of lung

3=disperses capsule, and 10-50% lung contains more serious infected zone

The lung of 4=＞50% contains many thing kitchen ranges and infects serious zone

C=compares, and I represents the known compound of a structural formula I, and II represents the compounds of a structural formula II

Table VIII

Disease degree by histological grade

Number of animals example number 0.5 1234

Contrast 0001 712 C R ₂=H, X=O, n=5 4200 06 II R ₂=OCH ₃, X=O, n=3 7100 011 II R ₂=OCH ₃, X=O, n=4 3410 015 II R ₂=OCH ₃, X=O, n=5 0341 024 C R ₂=Cl, X=O, n=4 0123 225 C R ₂=Cl, X=O, n=5 00044

Table VIII (continuing)

Histological grade:

0.5=find capsule＜10 in per two cross sections

1=disperses capsule, 5% of＜lung

2=disperses capsule, 5-10% or little infection focus of lung

3=disperses capsule, and 10-50% lung contains more serious infected zone

The lung of 4=＞50% contains the serious zone of many focal infections

C=compares, and I represents the known compound of a structural formula I, and II represents the compounds of a structural formula II

Table I X

Disease degree by histological grade

Number of animals example number 0.5 1234---contrast 0006 212 C R ₂=H, X=O, n=5,0610 033 II R ₂=NH ₂, X=N, n=2 00431

—X—(CH ₂) _n—X—

R ₃ R ₂ R ₂ R ₃42 II R ₃＝CH ₃，R ₂＝H，X＝O，n＝5 0 0 2 3 341 II R ₃＝CH ₃，R ₂＝H，X＝O，n＝4 0 0 2 4 244 II R ₃＝H，R ₂＝OCH ₃，X＝O，n＝3 6 2 0 0 0

Table I X (continuing)

Histological grade:

0.5=find capsule＜10 in per two cross sections

1=disperses capsule, 5% of＜lung

2=disperses capsule, 5-10% or little infection focus of lung

3=disperses capsule, and 10-50% lung contains more serious infected zone

The lung of 4=＞50% contains the serious zone of many focal infections

C=compares, and I represents the known compound of a structural formula I, and II represents the compounds of a structural formula II

Table X

Disease degree by histological grade

Number of animals example number 0.5 1234---contrasting 00125---35 II R ₂=NO ₂, X=N, n=3 00026 5mg/Kg36 II R ₂=NH ₂, X=N, n=3 14200 10mg/Kg37 II R ₂=NH ₂, X=N, n=4 04310 10mg/Kg38 II R ₂=NO ₃, X=O, n=4 00314 2.5mg/Kg

—X—(CH ₂) _n—X—

R ₃ R ₂ R ₂ R ₃40 II?R ₂＝H，R ₃＝H，X＝O，n＝4 1 2 3 1 0 10mg/Kg43 II?R ₃＝H，R ₂＝OCH ₃，X＝O，n＝51 3 1 2 0 5mg/Kg

Table X (continuing)

Histological grade:

0.5=find capsule＜10 in per two cross sections

1=disperses capsule, 5% of＜lung

2=disperses capsule, 5-10% or little infection focus of lung

3=disperses capsule, and 10-50% lung contains more serious infected zone

The lung of 4=＞50% contains the serious zone of many focal infections

C=compares, and I represents the known compound of a structural formula I, and II represents the compounds of a structural formula II

Table X I

Disease degree by histological grade

Number of animals

0.5 1234 contrasts, 00255 five amidine 10mg/Kg, 35210 fourth amidine 10mg/Kg, 10 20005 amidine 1mg/Kg, 10353 fourth amidine 1mg/Kg, 01353 five amidine 0.1mg/Kg, 00075 fourth amidine 0.1mg/Kg 00452

Table X I (continuing)

Histological grade:

0.5=find capsule＜10 in per two cross sections

1=disperses capsule, 5% of＜lung

2=disperses capsule, 5-10% or little infection focus of lung

3=disperses capsule, and 10-50% lung contains more serious infected zone

The lung of 4=＞50% contains the serious zone of many focal infections

Table X II

The number of animals of disease degree (bonded) the compound number rank groups by histological grade

0.5 not test-undissolved 3. 4400 04. 3410 05. 2410 06. in animal of test-undissolved 2. in animal of 1234 salt molten 129 25 351.^c7100 07. 5120 08. 3100 09. 8000 010. 2510 011. ^c3410 012. (five amidines) 20 26 12 2 013. ^c0242 014. 85110 15. ^d2471 016. not tests-undissolved 17. 1231 018. in animal ^c1302 119. 32300

Table X II (continuing)

The number of animals of compound number rank groups

0.5 123 420. 3310 021. ^e4110 022. ^c0143 023. 3120 124. ^d0123 225. ^d0004 426. 2141 027. 2113 028. 4362 129. 1412 030. 5300 031. test-synthesizing 32. test-not synthesizing 33. 0. 043 134. do not test 35. in animal in animal in animal^c0002 636. 1420 037. 0431 038. ^d0031 439. not tests 40. 12310 in animal

Table X II (continuing)

The number of animals of compound number rank groups

0.5 1 2 3 441. ^d 0 0 2 4 242. ^d 0 0 2 3 343. ^c 1 3 1 2 044. ^d 6 2 0 0 0

Histological grade:

0.5=find capsule＜10 in per two cross sections

1=disperses capsule, 5% of＜lung

2=disperses capsule, 5-10% or little infection focus of lung

3=disperses capsule, and 10-50% lung contains more serious infected zone

The lung of 4=＞50% contains the serious regional b. of many focal infections unless otherwise indicated, and all compounds are all tested under the 1.25mg/Kg condition at test e. under the 2.5mg/Kg condition at test d. under the 5mg/Kg condition at test c. under the 10mg/Kg condition

Table X III

The toxic chemical number effect 1 of amidine test-does not dissolve 2 in animal test-not dissolve 3 nothings 4 in injection site mild swelling 5 nothings 6 nothings-only test 7 nothings 8 tail end necrosis under the 5mg/Kg condition in animal, chronic poisoning, there were 3 dead 9 nothings 10 not have 11 down to the 7th day in the 10mg/Kg condition and do not have-only under the 5mg/Kg condition, test 12 some ypotension, tail oedema 13 acute poisonings, dead 14 do not have 15 severe hypotensions under the 10mg/Kg condition, acute poisoning, dead 16 test-do not dissolve 17 chronic poisonings in animal under the 10mg/Kg condition, to the 113rd day 2 dead 18 acute poisonings, dead under the 10mg/Kg condition under the 10mg/Kg condition

Table X III (continuing)

Compound number effect 19 20 some ypotension of shaking, the tail oedema, stronger anticoagulant effect 21 acute poisonings, 10,5 and the 2.5mg/Kg condition under dead 22 acute poisonings, spasm 24 does not have during dead 23 fast injection under the 10mg/Kg condition, because testing 25 under the 2.5mg/Kg condition, solubleness do not have, since solubleness test 26 under the 2.5mg/Kg condition do not have 27 do not have 28 in the injection site the slight oedema 30 of slight swelling 29 tails do not have 31 do not test 32 do not test 33 some anticoagulant effect 34 do not test 35 since solubleness under the 5mg/Kg condition, test, nontoxic 36 do not have 37 nothings 38 acute poisoning under 10mg/Kg and 5mg/Kg condition, and test is not poisoned under the 2.5mg/Kg condition

Table X III (continuing)

Compound number effect 39 does not test 40 in animal do not have 41 owing to solubleness is tested under the 2.5mg/Kg condition, nontoxicity 42 is acute poisoning under 10mg/Kg and 5mg/Kg condition, under the 2.5mg/Kg condition test do not poison 43 under the 10mg/Kg condition acute poisoning, " little trembling " and arrhythmia, do not poison 44 because solubleness is tested nontoxicity under the 2.5mg/Kg condition in test under the 5mg/Kg condition

Table X IV

The Lan Shi giardia lamblia with to amidine Instance number X n R ₂IC ₅₀(uM) 45 O, 2 NH ₂1183.846 N 2 NH ₂743.747 O 3 H 12.348 O 3 NH ₂15.449 O 3 OCH ₃3.250 N 3 H 68.251 N 3 NO ₂84.252 O 4 H 61.653 O 4 NH ₂96.854 O 4 NO ₂53.2

Table X IV (continuous table)

The Lan Shi giardia lamblia with to amidine

R ₁, R ₃=H instance number X n R ₂IC ₅₀(uM) 55 O, 4 OCH ₃68.856 O 4 Cl 27.757 N 4 H 50.758 N 4 NH ₂107.759 O 5 H 76.560 O 5 NH ₂66.361 O 5 NO ₂26.162 O 5 OCH ₃24.063 O 5 Cl 16.364 N 5 H 40.665 N 5 NH ₂48.666 N 5 NO ₂29.967 O 6 H 34.368 O 6 NH ₂38.769 N 6 H 4.270 N 6 NH ₂21.9

Table X V

A Lan Shi giardia lamblia and an amidine

Instance number X n R ₂IC ₅₀(uM) 71 O, 3 H, 560.172 O, 4 H, 105.873 O, 5 H, 55.874 O, 6 H 84.2

Table X VI

The Lan Shi giardia lamblia with to tetrahydroglyoxaline Instance number X n R ₃R ₂IC ₅₀(uM) 75 O, 3 H OCH ₃25.476 O 4 H H 26.077 O 4 CH ₃H 346.378 O 5 H H 39.279 O 5 CH ₃H 373.380 O 5 H OCH ₃47.1

Table X VII

The current compound that is used for the treatment of giardiasis of the U.S.

And use ³The H thymidine mixes the IC that measures them ₅₀

Metronidazole plain BP.98 99 _1eThe Nifurazolidone Quinacrime _.HClIC ₅₀=3.65 IC ₅₀=1.90

IC ₅₀=1.41

IC ₅₀=0.91 μ M_ IC ₅₀=0.7 μ M_ IC ₅₀=1.18 μ M_IC ₅₀=2.14 μ M § IC ₅₀=0.60 μ M § IC ₅₀=1.09 μ M § IC ₅₀=1.08

IC ₅₀=2.50 IC ₅₀=1.59

IC ₅₀=2.21 μ M ± IC ₅₀=0.43 μ M ± IC ₅₀=4.02 μ M ±

Original seed-MB (ATCC30957) _ original seed-BRIS/83/HEPU/106 (Boreham et al., 1984) § original seed-BRIS/82/HEPU/4l (Boreham et al., 1984)

Original seed-BRIS/83/HEPU/120 (Boreham et al., 1984) ± original seed-BRI/83/HEPU/136 (Boreham et al., 1984)

Table X VIII

Leishmania mexicana with to amidine

R ₁，R ₃＝H

(new) (existing) instance number X n R ₂IC ₅₀Grade IC ₅₀Grade

(uM) (uM)81 O 3 OCH ₃ 1.69 10 1.93 1282 N 4 H 0.58 3 0.60 383 O 4 H 1.23 7 1.19 984 O 5 H 0.65 4 0.75Ave. 585 O 5 NO ₂ 2.57 13 1.07 886 O 5 NH ₂ 2.32 12 0.95 687 O 5 OCH ₃ 6.53 16 2.12 1388 N 5 H 0.35 2 0.35 189 O 6 H 0.33 1 0.43 290 O 5 Cl 1.11 6 1.06 7

Table X IX

A leishmania mexicana and an amidine R ₁, R ₃=H

(new) (existing) instance number X n R ₂IC ₅₀Grade IC ₅₀Grade

(uM) (of18) (uM) (of18)91 O 3 H 7.20 17 ＞4.5 17-1492 O 4 H 3.28 15 ＞4.5 17-1493 O 5 H 1.66 9 1.51 1194 O 6 H 2.92 14 1.35 10

Table X X

Leishmania mexicana with to tetrahydroglyoxaline

(new) (existing) instance number X n R ₃R ₂IC ₅₀Grade IC ₅₀Grade

(uM) (uM)95 O 5 H H 1.06 5 0.67 496 O 5 CH ₃?H 12.2 18 ＞4.5 17-1497 O 3 H OCH ₃ 1.71 11 ＞44.5 17-1498 O 3 H H 1.47 8 —— X

Table X XI

Plasmodium falciparum with to amidine

R ₁, R ₃=H

^W2Grade ^D6Grade instance number X n R ₂IC ₅₀(uM) (of18) IC ₅₀(uM) (of16) 99 O, 3 OCH ₃0.065 6 0.077 11100 N, 4 H, 0.057 3 0.025 3/4101 O, 4 H, 0.120 13 0.057 8102 O, 5 H, 0.083 11 0.038 5103 O, 5 NO ₂0.069 8 0.071 10104 O, 5 NH ₂0.076 9 0.046 6/7105 O, 5 OCH ₃0.066 7 0.106 14106 N, 5 H, 0.022 1＜0.020 1107 O, 6 H, 0.345 17 0.118 16108 O, 5 Cl 0.165 16 0.107 15

Table X XII

A plasmodium falciparum and an amidine

R ₁，R ₃＝H

^W2Grade ^D6Grade instance number X n R ₂IC ₅₀(uM) (of18) IC ₅₀(uM) (of18) 109 O, 3 H, 0.156 15 0.194 18110 O, 4 H, 0.043 2 0.046 6/7111 O, 5 H, 0.080 10 0.069 9112 O, 6 H 0.125 14 0.132 17

Table X XIII

Plasmodium falciparum with to tetrahydroglyoxaline

^W2Grade ^D6Grade instance number X n R ₃R ₂IC ₅₀(μ M) be IC (of18) ₅₀(uM) (of18) 113 O, 5 H H, 0.104 12 0.023 2114 O, 5 CH ₃H 1.817 18 0.089 12115 O 3 H OCH ₃0.060 5/4 0.100 13115 O, 3 H H 0.060 5/4 0.025 3/4

Claims (1)

1. be prepared as follows the method for the compound of formula

N=2-5 in the formula, R ₂Represent H ,-OCH ₃, No ₂Or NH ₂, and the Am representative:

Each R in the formula ₁Be H or two R ₁The group representative-(CH that on same miaow base, combines ₂) _m-, m=2 in the formula, 3, or 4, and R ₃Be H ,-CH ₃, or-CH ₂-CH ₃,

This method comprises

(1) method A:

Work as R ₂Be-OCH ₃, when n=2-5 and Am represent following formula,

R in the formula ₁Be H or two R ₁The group representative-(CH that on same miaow base, combines ₂) _m-, m=3 or 4 in the formula, and R ₃Be H ,-CH ₃, or-CH ₂-CH ₃

Will as shown in the formula the cyanophenol that is replaced by methoxyl group

Use as shown in the formula dibromo alkane carry out alkylation

Br-(CH ₂) _n-Br

In the formula definition of n as mentioned above with form as shown in the formula the cyano group intermediate compound

R in the formula ₂With the definition of n as mentioned above and subsequently cyano group is changed into the miaow base with generate as shown in the formula required product:

N in the formula, R ₁And R ₂Definition as mentioned above; Perhaps

(2) method B:

Work as R ₂Be NO ₂, n=2-5, and Am is when representing following formula

Each R in the formula ₁Be H or two R ₁The group representative-(CH that on same miaow base, combines ₂) _m-, m=2 in the formula, 3 or 4, and R ₃Be H ,-CH ₃, or-CH ₂-CH ₃Will as shown in the formula cyanophenol Use as shown in the formula dibromo alkane carry out alkylation

Br-(CH ₂) _n-Br

In the formula definition of n as mentioned above with form as shown in the formula the cyano group intermediate compound

The definition of n is carried out this cyano group intermediate compound nitration as mentioned above then to form dinitrobenzene dicyano compound as follows in the formula

The definition of n subsequently, changes into the miaow base with cyano compound as mentioned above in the formula, with generate as shown in the formula required product:

N in the formula, R ₁, and R ₃Definition as mentioned above; Perhaps (3) method C: work as R ₂Be NH ₂, when n=2-5 and Am represent following formula

Each R in the formula ₁Be H or two R ₁The group representative-(CH that on same miaow base, combines ₂) _m-, m=2 wherein, 3, or 4, and R ₃Be H ,-CH ₃, or-CH ₂CH ₃, with the cyanophenol of following formula:

Dibromo alkane with following formula carries out alkylation

Br-(CH ₂) _n-Br

In the formula definition of n as mentioned above with form as shown in the formula the cyano group intermediate compound:

The definition of n is nitrated with the cyano group intermediate then as mentioned above in the formula, with form as shown in the formula dinitrobenzene dicyano compound

The definition of n changes into amino with the nitro in the formula as mentioned above again in the formula, be able to as shown in the formula compound:

The definition of n changes into the miaow base with the cyano group in the formula as mentioned above and subsequently again in the formula, with generate as shown in the formula required compound

N in the formula, R ₁And R ₃Definition as mentioned above; Perhaps (4) method D: work as R ₂Be H or-OCH ₃, n=3 or 4, and Am is when representing following formula

Two R in the formula ₁The group representative-(CH that on same miaow base, combines ₂) ₂-, and R ₃Be H; Or CH ₃, will as shown in the formula cyanophenol

R in the formula ₂Definition use as mentioned above as shown in the formula dibromo alkane carry out alkylation

Br-(CH ₂) _n-Br

In the formula definition of n as mentioned above with form as shown in the formula the cyano group intermediate compound

N and R in the formula ₂Definition make as mentioned above and subsequently the cyano group intermediate with as shown in the formula compound reaction

NH ₂-(CH ₂) ₂-NHR ₃

R in the formula ₃Be H or-CH ₃So that the cyano group in the formula is changed into imidazolyl, to generate required following formula: compound R in the formula ₂, R ₃With the definition of n as mentioned above.

Images