JPS60184018A - Antiprotozoal agent - Google Patents

Abstract

PURPOSE:To provide an antiprotozoal agent containing d'-deoxyinosine as an active component, exhibiting high selective activity to the pathogenic protozoa of leishmania and trypanosoniasis at a low concentration, and having low side effect. CONSTITUTION:The objective antiprotozoal agent contains d'-deoxyinosine as an active component. It has been found that d'-deoxyinosine is remarkably effective to the pathogenic protozoa of leishmania and trypanosomiasis in high selectivity at a low concentration. Furthermore, it has low side effect, and is useful for the prevention and remedy for leishmania and trypanosomiasis. It is administered orally or parenterally in the form of powder, tablet, injection, etc. using conventional carrier, vehicle, diluent, etc. Dose; 5-100mg of d'-deoxyinosine per kg daily for oral administration and 1-40mg/kg daily for parenteral administration.

Description

DETAILED DESCRIPTION OF THE INVENTION The present invention relates to an antiprotozoal agent. More particularly, the invention particularly relates to Leishmania Ir.
, +1. )Tripanosoma
This invention relates to antiprotozoal agents for use in the prevention and treatment of diseases caused by eugenic protozoa belonging to the genus.

Leishmaniasie and Trypanosomi
asie) are diseases caused by parasitic protozoa belonging to the genus Leishmania and Trypanosoma, respectively, and are prevalent in specific regions from temperate to tropical regions.

Examples of parasitic protozoa belonging to the genus Leishmania include Loishmania donoba, which is the one-billionth protozoan of KAla-azar.
va, ni) + oriental axis (oriental 5ore)
Leishmania tropic
a) Also, the pathogenic protozoan of Eepundia in South America, Leishmania gwojiriensii)
iehmania braziliensi8) and Leishmania mexicana (Leishmania m
exicana), etc. are known. In addition, examples of parasitic protozoa belonging to the genus Trypanosoma include African trypanosomiasis (African Trypanosomiasis).
omia8is) embryonic protozoan Trypanosoma gambiens (I'rypanosoma gambienee)
+) Libanosoma rhodesiens (Trypan)
osoma rhodesiense), but also American trypanosomiasis (AmericantrypFL).
Trypanosoma spp.
Trypanosoma cruzi and the like are known.

Pentamidine is currently used as a therapeutic agent for diseases caused by these parasitic protozoa.
amidine) + bent stun (pentosta)
ne) Also amphotericin B (amphotericin B)
ine B) suramin + stilbamine isethionate
onate ) + ) Ribalsamide (TrypPLre
amide), but all of these drugs have strong side effects, so patients need to be hospitalized and receive these drugs under the strict supervision of a doctor, as well as undergo treatment.

However, there are relatively many developing countries in which Finnemaniasis and trypanosomiasis are endemic, and these countries lack skilled medical practitioners. There is a strong desire to develop an anti-parasitic protozoan agent that can be used against 113 stools.

On the other hand, 3'-deoxyinosine is an already known compound
S [Pretan of the Chemical Society]
Op Japan (Bull, Chem, Soc.

Jpn) Vol. 43, No. 3922 (1970)], however, little is known about the biological effects of this compound, and nothing is known about its anti-fishmanial and anti-trypanosome activities.

The inventors of the present invention have conducted intensive research in order to develop a drug that has few side effects and is effective for the treatment of diseases caused by pathogenic protozoa such as phyciemaniasis and trypanosomiasis. We have discovered that deoxyinosine is highly selective and highly effective against the pathogenic protozoa of Finnemaniasis and trypanosomiasis, and based on this finding, we have completed the present invention.

That is, the present invention provides an antiprotozoal agent containing 3'-deoxyinosine.

3'-Deoxyinosine is described, for example, in the above-mentioned literature [Bull, Chem, 8oc, Jpn.
M43, page 3922 (1970)] can also be produced by the method described in Reference Examples below.

3'-deoxyinosine tl - When administered to humans or other warm-blooded animals for the prevention or treatment of diseases caused by the above-mentioned pathogenic protozoa, it can be administered as such or by commonly used methods, such as pharmacologically. l “Acceptable carriers, excipients,
By using diluents, solubilizing agents, etc., you can prepare powder, lI! I'1 tablet + 'dt tablet, capsule.

It can be administered orally or parenterally in the form of injections, suppositories, etc. The dosage varies depending on the symptoms, the nutritional status of the affected animal, the route of drug administration, etc., but for example, R1! When administered orally for the treatment of affected adults, it is administered approximately 1 to 5 times a day, approximately 5 to 100 W/day as 3'-deoxyinosine, and once per day when administered parenterally. ~5 times, IJ I~40M per day
9 body weight is preferably used for f/.

3′-deoxyinosine is used in the examples described below.
It has been confirmed that it exhibits its antiprotozoal activity at low concentrations. In addition, it has also been confirmed that animal tissues have significantly lower Wj properties, and the concentration ratio between them is 25.
It reaches more than 0 times. This indicates that the compound of the present invention has a highly selective activity against protozoa, 3'-deoxyinosinca 1jtJe 1tFj.

Next, the details of the present invention will be explained using reference examples and W examples.
The present invention is not necessarily limited to these.

Reference Example 3 - Deoxyadenosine (85011'f/) and 7 denosine deaminase (354, Sigma A 9876) were added to a 100 mM bicarbonate-triethylamine solution (90 g).
? , pH 7.1), and the reaction was carried out at 37°C for 4 hours. The reaction solution was evaporated to dryness under reduced pressure at 30°C to obtain 680W crystals of 3'-deoxyinosine in ethanol.

Example 1 (1) Culture was carried out at 25°C using 3'-deoxyinosine proliferation serum for Leishmania tropica + 25mM Hepes chloride (containing hemin and hoxanthine). 1×
Cell density glue of 105 cells/ml, tropica
3'-deoxyinosine was added to the medium containing f at a final concentration of 9 × f (r5M -3 × 10 M tonal-like (1f2M111f)), and 3'-deoxyinosine was added to the medium without treatment (with the group The cells were cultured at 25° C. for 48 hours, and the degree of total cell nephropathy was measured.Proliferation % was calculated using the following formula.

N4J cell quality of the 3'-deoxyinosine addition group (cell density after 48 hours - cell density at the start of the experiment) IM degree (1411 cell abundance after 48 hours - cell density at the start of the experiment) control)
If EC5o is the concentration of 3'-deoxyinosine that inhibits proliferation by 50% compared to L.

E of 3'-deoxyinosine for P. tropica
C5° was 4.43xlOM (average value of two dormitory songs).

(2) The EC5o of 3'-deoxyinosine for milk 1NII cells (FM3Am derived from mouse breast cancer)
This was done using cells. ES medium containing 1M3A cells at a cell density of 5 x 10 m cells/wl in 2% beef tallow serum).
'-Deoxyinosine t-Mk final concentration 1.2X10-'
M - 3 x 10 M and cultured at 37C (5% CO
2) I did. EC50 was calculated in the same manner as for L. troplca. 3'-aligned with FM3A vacuole
The EC5o for deoxyinosine was 1.25X10M (average of two experiments).

3 for L, tropica and FM3AMll follicles.
’-deoxyinosine “C50 value ratio (L, trop
ica.

The EC value/EC5o value of hM3Ailtl cells was approximately 0.0035, indicating strong selection against protozoan parasites.

The results of this example are summarized in FIG. 1.

Example 2 Amastigotes collected from the pancreas of infected hamsters) (
amastigote) were added to male mice (B
Leishmania donovai (Leishmania donovai)
ni ) times for 5 consecutive days. On the 7th day after the first administration of the drug, the mice were sacrificed, the hepatic fistula was excised, and after weighing, a compression cord was prepared from the liver incision. After staining this product with Giemsa, the number of amastigotes per 500 hepatic axon nuclei was counted using Gu Huanjing. Calculate the total number of parasites by
Protodology (J.

Protozoology) Volume 5, Page 269 (19
The average value of 5 animals in each group was calculated using the method described in 1958).

The results obtained in this example are shown in Table 1 in terms of inhibition rate.

Table 1 Example 3 3'-Deoxyinosine 100 ft - Distilled water for injection 50
Disperse at 0vtl and add WI soda to pHt-about 8
Adjust to -9. Add distilled water for injection to make 11.

Dispense 5*/ each into injection containers using aseptic techniques to prepare injections.

Example 4 (1) 3'-deoxyinosine 5(R(2) Lactose 30.7f (3) Starch 1i (4) Hydroxypropyμcellulose 2F(5)
Magnesium stearate 0.3F100.0f (i) * Mix (2) and (3), granulate with (4) as a binder, add (5) and press into tablets, each tablet weighs 10011v. Manufacture 1000 tablets.

[Brief explanation of drawings]

Figure 1 shows 3'-deoxyinosine glue, tropic
The results for a and pine 1f are shown. Curve A and B beams. tropica
+Curves C and D are 3'- to mouse FM3AM follicles.
A deoxyinosine concentration-proliferation curve is shown.

Claims (1)

[Claims] Antiprotozoal agent containing 3'-deoxyinosine