JPS601129A - Antipiroplasmic agent - Google Patents

Abstract

PURPOSE:To provide a drug containing at least one kind of tetrocarcin compound and effective as an antipiroplasmic agent and antimalarial agent. CONSTITUTION:The object agent contains at least one of the tetrocarcin compounds of formula. Piroplasmosis is a nematodosis caused by piroplasmas of wide sense including the nematodes of Babesia family and Theileria family, carried by mites, and exhibits acute or chronic principal symptoms of pyrexia, anemia, jaundice, blood urine, etc. The tetrocarcine compound of formula is useful not only as an antipiroplasmic agent for animals, especially cattle, but also as an antimalarial agent for man and other animals (e.g. poultry). Dose: 0.1- 20.0mg/kg, once a day, 1-7 doses a cure, continuously or intermittently for antipiroplasmic agent, and 0.5-10mg/kg in the same manner as above for antimalarial agent.

Description

DETAILED DESCRIPTION OF THE INVENTION The present invention relates to an anti-pyroplasma or anti-malarial agent containing at least one member of the tetrocardin class and effective for the prevention and treatment of piroplasmosis or malaria.

Piroplasmosis is a protozoal disease caused by piroplasma, which is a combination of protozoa of the Babesiaceae and Theileriaceae families, and is an acute or chronic disease transmitted by ticks and mainly characterized by fever, anemia, jaundice, and bloody urea. It is.

Various treatments have been tried since ancient times, but it is extremely difficult to prevent, and it frequently occurs in various parts of the world, causing serious economic losses. On the other hand, in Japan as well, it occurs frequently in pasture-raised cattle, and in recent years it has also been increasing in caged cattle, making it a problem.

As a drug, there are 8-aminoquinoline derivatives, but recently, many resistant strains have appeared, and the current situation is that they have little practical value in terms of effectiveness, toxicity, etc.

Therefore, the present inventors investigated the small animal parasitic Babesia rodt+ain+ (hereinafter referred to as B
As a result of extensive research into effective anti-pyroplasma agents using R. Furthermore, Theileria sergeenti (Tt+
eileria sergenti) (hereinafter referred to as TS) (including drug-resistant strains) was infected to harvested cattle and its anti-pyroplasma effect was confirmed, and its practical value was also revealed. Ta.

Tetrocardins are a group of antibiotics filed by the applicant and have the following structure.

Te C1 Tetrocardin A: The above compound Tetrocardin B: H tetrocardin instead of Sugar D? : H tetrocardins 1 in place of sugars C and D: 11 tetrocardins E2 in place of sugar BD:
Acetyl instead of sugar B~D, 110-5- instead of C113 0- in A ring Trocardin F-1: H instead of sugar A-D Tetrocardins-2: instead of sugar A-'D H.

11 instead of sugar E Tetrocardin G: C1120 instead of C110 at position 23
+1 tetrocardin H: GO instead of Cll0 at position 23
2+1 In addition to the above, there are tetrocardins C, D, I, J, L, F, i, etc. having similar structures.

Also included are compounds in which the 9.21st position of these compounds is an acyl group (diazylate) and those in which the 9,17.21st position is an acyl group (triacylate). The applications for these tetrocardins are as follows, but the name tetrocardin is not used, but DC-11-A, DC-11-B, etc.D
There are cases where the name C-11 is attached: JP-A-54-
138501 (Special Publication No. 56-38159), Japanese Patent Publication No. 1973
5-79322.56-139500. U.S. Special I1R No. 4,346,075 (tetrocardin A).

JP 56-1159794.56-122392 (hereinafter referred to as "Nitetrocalcin B"), 'IHIJ 1987-75
500.56-12239'2 (Tetrocardin C), JP-A-56-122392 (Tetrocardin D)
), JP-A-57-38796 (Nato 1 Cocalcin E1
．． E2) + JP-A-57-53498 (tetrocardin F, G, H), JP-A-57-171997 (tetrocardin I, J, ni, L).

M), Il & JP 57-7455 (tetrocardin F-1,1,'-2), JP 57-7479 (diazylate, triacylate tetrocardin, as mentioned above, is suitable for animals, especially cattle) nm as an anti-pyroplasma agent, but further research has revealed that C is also useful as an anti-malarial agent for humans or animals (for example, domestic animals).

Tetrocardin buccal may be administered to humans or other animals orally or parenterally. Zunawaji, Note 1. In the case of J, it may be dissolved directly in water or physiological saline, or an antioxidant (sthorium pyrosulfite, etc.).

Soothing agents (procaine hydrochloride, etc.), preservatives (methyl paraoxybenzoate, propyl paraoxybenzoate, etc.), pH
A regulator (sodium hydroxide, etc.) or the like may be further added.

In addition, tetrocardins contain diluents (for example, starch, silosaccharide, lactose, calcium carbonate, kaolin, etc.), fillers (for example, lactose, starch, calcium carbonate, calcium phosphate, kaolin, bentonand, talc, etc.), lubricants ( For example, powders, tablets, and granules can be prepared by adding pharmaceutical ingredients such as stearic acid, paraffin, boric acid, silica, sodium benzoate, polyethylene glycol, etc.).

It can also be administered in the form of capsules, suppositories, suspensions, emulsions, etc.

When tetrocardins are used as anti-pyroplasma agents, administration it 0. 1-2 0. Onv/
kg (especially 0.32 to 9.6 ■/kg) (as a tetrocardin), the number of administrations was 1 to 7 in 181 doses.
One course is defined as one course, and the drug is administered continuously or intermittently. The common method of administration is intravenous injection, but subcutaneous administration. Intramuscular injection,
Intraperitoneal administration and oral administration are also possible.

When tetrocardins are used as an antimalarial agent, the dosage may be 0.5 to 10 kg/kg (as tetrocardins), and the number of administrations and method of administration may be the same as above.

Aspects of the present invention will be illustrated below by way of examples.

Example 1.

Blood was collected from BR-infected mice, and the number of parasites was reduced to 8.
Infected blood cells were prepared at 6 x 105 cells/0.2 ml and inoculated to 5 mice per group i and p to form infected mice.

Tetrocardin A and B were subcutaneously injected per animal once a day for 7 consecutive days starting at the same time as infection in all groups. Tetrocardin A
The dosage is 0.1ml each as shown in Table 1,
Tetrocardin B is 0.1 tw/0. The In+1 solution was made into 0.1 ml each.

Mouse survival rates are shown in Table 1.

Of the five animals in the non-medicated control group, one died on the 7th day and 4 on the 8th day, and autopsy revealed marked anemia and enlarged liver.

Anemia, enlarged tiles, and enlarged mesenteric lymph nodes were evident, and jaundice was sometimes evident. BR parasitism is highly observed within red blood cells.

On the other hand, 0.025■/day/mouse to tetrocardin and B'R to tetrocardin B061■/day/mouse.
If the drug is administered for 7 consecutive days from the day of infection, all cases survive for more than a month and are effective.

Table 1 Example 2 Tests were conducted using infected mice produced in the same manner as in Example 1. Tetrocardin 8 was administered at the dosage shown in Table 2, and the number of subcutaneous administrations was shortened to 1 to 5 times. The results are shown in Table 2.

All five animals in the non-medicated control group died by the 9th day, and the m-examination findings and protozoan detection results were the same as in the control group of Example 1.

Table 2 Example 3 Tests were carried out in the same manner as in Example 1. Dosing is 0.1
ml, the dosing schedule is shown in Table 3 Table 1-m3 *The day of infection is day 0.

Blood smear test: Blood smears were prepared for some patients during the course of the treatment and tested for the presence or absence of BR infection.

Challenge: In groups that survived more than 1 month after infection, some mice (e.g., 3 out of 5 in the all-survival group) were challenged by intraperitoneal inoculation with the same number of BRs as at the time of inoculation.

Serological response: Mice that survived the challenge, along with surviving mice that were not challenged, were challenged f&
The animals were sacrificed after more than one strain had elapsed, and plasma was collected. This plasma and Parasitemia
An agar gel precipitation reaction was performed between the antigen and the plasma antigen collected at the peak stage (a state in which protozoa appear in red blood cells).

The results are shown in Table 4.

0.4■, and 0.2 in the single-dose group. and 4th day dosing, 0
, 2■, θ~1, 2~3.4~5 and 6~7 in the 2nd group
Dosing daily, 0.1■; in the 3-dose group, dosing daily for 4 to 6 days, 0
．． After the initial infection, all cases survived and were effective after 7 doses administered in 05■.

Considering the presence or absence of barasithemia during the course of infection, the results of challenges to surviving cases, and the results of sedimentation reaction of serum in agar gel at the time of final sacrifice, it was found that barasithemia during the course of infection was detected in the shade 11J. H. The reason why all the mice died after the challenge was in groups ① and 0. It was revealed that administration of 0.4 mg once or twice at 0.21 mg iJ at the early stage of infection was extremely effective.

Example 4 As a result of a test using BR parasitic on small animals, it was revealed that tetrocardin A and TeI/localcin B were very effective. We investigated whether it is effective and practical.

TS on a cow weighing around 200 kg with +1'll eviscerated
TSS by subcutaneously inoculating 109 Fukushima strains once.
It looked infected.

All medications are administered by intravenous injection, and the dosage is 0 per animal.
．． 32■/kg and 3.2■/kg, the number of doses was 2
~3 times on consecutive days or every other day. The results are shown in Figures 1-3.

0.32+w/kg/day/cow, TSO) When the parasitism rate exceeded 25%, the drug was administered once a day for 3 consecutive days.

When administering 3.2 kg/day/cow twice, the dose was administered once when the TS parasitism rate exceeded 23%. Since the parasitic rate was 3.5% on the 7th day of administration, the same amount was administered once more, and a considerable decrease of 0.1% was observed on the 20th day.

3.2 mg/kg/day/3 times every other day for cows, TS
Administration was started when the parasitism rate exceeded 22.5%. The parasitism rate rapidly decreased to 0.1 on the 4th day after the end of treatment.
%, which decreased to 0% on the 6th day, indicating that it is effective.

Example 5 A TSS infection color was produced in the same manner as in Example 4, and the efficacy of tetrocardin A was investigated.

The dosage of tetrocardin A is 6.4 / 1111
kg was administered intravenously 2-3 times every other day. The results are shown in figure ff14°5.

In the case of dosing twice every other day at 6.4 μ/kg/day for cows, dosing was started when the TS parasitism rate exceeded 14%.

The parasitism rate decreased rapidly, and 1.
0%, and 0% on the 6th day, indicating that it is effective.

G, 4nqr/+r/day/Cows administered 3 times every other day'rS (7) Dosing was started when the parasitic rate exceeded 20%.

The parasitism rate was already 0.6% on the day of the end of treatment, and it was 0-0 on the 2nd day.
+1596% It becomes 0% on the third day, and it is extremely
) Anti-viroplasmic effects were observed. Even after that, the parasitism rate was 0.
% continued for more than 7 weeks, indicating that the anti-pyroplasma effect was sustained.

Example 6゜1'ss dyed beef was prepared in the same manner as in Example 4, and tetrocardin 8, commercially available anti-pyroplasma agents, diminazine acetate preparation (Ganazesoku) and 8-aminoquinoline preparation (Bamaquin) were used. ) was conducted.

The dosage for tetrocardin is 6.4■/kg per animal.
Ganazesoku is administered intravenously twice every other day, and 10.
0■/kgi! [! Intramuscular pamaquine was injected subcutaneously at 1.6 kg/kg for 2 consecutive days.

Medication was carried out when the TS parasitism rate of each cow exceeded 20%. As a result, the parasitic rate of cows treated with tetrocardin A rapidly decreased to 1.5% on the 4th day and 0% on the 7th day, indicating that the treatment was effective.

On the other hand, in the cows treated with Ganazesoku, the parasitic rate decreased to 5% on the 6th day, but it hardly decreased after that and increased to 15% after 14 days, showing no effect.

In the cows treated with Pamaquin, the parasitic rate decreased to 10% on the 6th day, but it hardly decreased after that and after 140 days, an increasing tendency was observed, and there was no effect.

Example 7 A TS-infected cow was produced in the same manner as in Example 4, and the efficacy of tetrocardin A upon subcutaneous injection was examined.

Dosage for tetrocardin is 3.2■/kg and 6,
The dosage was 4 mg/kg twice every other day, and administration was started when the parasitism rate exceeded 25%.

When 3.2 kg/kg was administered twice every other day, it became 0.2% on the 7th day after completion of administration, which was clearly shown to be effective.

When administering 6.2■/kg twice every other day, it was 1.2% on the 3rd day after completion of administration and 0% on the 60th day, indicating that it is as effective as intravenous administration. . Furthermore, no side effects such as induration, edema, and fever were observed during administration.

Example 8 Safety of cattle with tetrocardin A administration per cow:
Even when administered intravenously, subcutaneously, intramuscularly, intraperitoneally, or orally once a day for 2 to 7 consecutive days at 0.32■/kg to 20.0■/kg, fever, anorexia, vomiting, pain, induration, and edema occurred. No side effects were observed from clinical findings or blood test findings.

Example 9゜Plasmodium bergei (Plasmodium
bergbei) (hereinafter referred to as PB) was collected from an infected mouse and diluted with physiological saline until the number of protozoa was approximately 2 x 106/10.2
Infected blood cells were prepared in a volume of 1.5 ml, and infected mice were inoculated i and p into 5 mice per group. Tetrocardin Δ was administered once per day per animal from the same time as infection in all groups7
The patient was injected subcutaneously for 0 days, and the progress was observed. Dosages for tetrocardin were 0.025, 0.05 and 0.1 /mouse/day.

As a result, of the five animals in the non-medicated control group, two died on the gold side on the seventh day, two on the eighth day, and one on the ninth day, and infected blood cells were found on the gold side at autopsy.

On the other hand, when the tetrocardin Δ group was administered at 0.025■/mouse/mouth for 7 consecutive days, one mouse died on the 8th day, 2 on the 10U, and 2 on the 11U, and the gold side died. There was not much difference between the control area and the control area.

0.05■/mouse/day for 7 consecutive days, the 16th U.
One animal each died on the 19th, 20th, and 25th day, but one animal survived beyond the 40 U, and a slight effect was observed.

In the group treated with 0.1 μ/mouse/day for 7 consecutive days, no symptoms were observed on the gold side, and the drug survived for 40 days or more, proving to be effective. Blood was collected from the tail vein of the mice in this group at 15 U and tested for malaria parasites, but the result was negative on the gold side. In addition, the animal was sacrificed on the 44th day, but no infected blood cells were found on the gold side, and no significant changes were observed at autopsy, and no malaria parasites were found in smears of the liver, tiles, lungs, kidneys, heart, thymus, or brain.

So, we pooled the above 7 organs from 5 animals and made saline solution! A suspension was prepared and two mice were inoculated i and p into second generation mice. 2
Since both of the mice in the second generation did not develop any symptoms, they were sacrificed on the 36th day after inoculation. No significant changes were found at autopsy; liver and Il.
No malaria parasites were found in organ smears of the lungs, kidneys, heart, or thymus, demonstrating the effectiveness of the drug.

Example 10 A test was conducted using PB-infected mice produced in the same manner as in Example 9.

The dosage of tetrocardin A is 0.000 mg per animal. Engineering.

The doses were 0.2 and 0.4 μ/mouse/day, and the frequency of subcutaneous administration was once a day for 7 consecutive days.

As a result, all animals in the no-medication control group died by the 8th day, but those in the group treated with TEI/localcin A at 0.1 μ/mouse/day or more survived without developing symptoms. 4
It was euthanized on day 0 and day 1. During this period, the test results for protozoa at the 15th U and at the time of sacrifice were negative, and the second generation mice, which were inoculated with the organ emulsion of the first generation mouse i and p at the time of sacrifice, passed without developing any disease and were sacrificed on day 40. Test results for protozoa were negative, indicating that the drug was effective.

Example 11 A test was conducted using 5 PB-infected mice produced in the same manner as in Example 9.

The dosage of tetrocardin B is 0. l.

(), 2 and 0.4q/mouse/day, and the number of subcutaneous administrations was once a day for 7 consecutive times. As a result, the no-medication control group 5
The animal died by the 81st day, and infected blood cells were found in the animal during autopsy.

On the other hand, in the tetrocardin B administration group, there was a slight increase in the number of surviving mice when the dose was 0.1+ng/mouse/l;
All five animals died, one each on the 1st day, 13th day, 17th day, 20th BIEL, and 1st day on the 23rd.

In the case of 0, 2 nv and 0.4 nv/mouse/day, the mice survived without developing any symptoms and were sacrificed on day 40. During this period, the test result for protozoa was negative on day 1 and at the time of sacrifice, and at the time of sacrifice, the organ emulsion of the first generation mouse was taken.
l. The second generation of inoculated mice passed without developing any disease.
I killed it on the 40th, but the protozoan test results were negative.
The drug was found to be effective.

[Brief explanation of the drawing]

Figures 1 to 5 show the effects of administering tetrocardin to 1゛S-infected culled cows. In Figures 1 to 5, the vertical axis shows the number of protozoan parasitized red blood cells per 1000 red blood cells, and the horizontal axis shows the number of days. Arrows indicate time points of administration, each one arrow ff
! Figure 1 shows the dose of 0.32■/kg, Figures 2 and 3 show the dose of 3
．． 2■/kg administration, 6.4 w/kg in Figures 4 and 5.
kg administration is shown. Patent Applicant (102) Kyowa Hakko Kogyo Co., Ltd. Procedural Amendment Written on May 7, 1982 by the Commissioner of the Patent Office 1. Indication of the case 1982 Patent Application No. 107406 2. Name of the invention: Anti-pyroplasma agent 3. Relationship with the case of the person making the amendment Patent applicant postal code 100 Address 6-1 Otemachi-chome, Chiyoda-ku, Tokyo Name
(102) Wakiwa Shoten Industry Co., Ltd. (TIEL: 03
-201-7211 Extension 2751) 5. Contents of correction Add the following example after line 3 on page 20. Example 12 A test was conducted in the same manner as in Example 3. Table 5 *1 Δ B C: Tetrocalcin Δ, B, C *2
Tetrocardin B and C had a considerable effect, and BR-infected mice survived on the gold side. Looking at the results after challenge, in the tetrocar 273 group, 1 out of 3 challenged mice
One of the mice died on the 8th day, but two survived, and the in-gel precipitation reaction of the two mice that were not challenged was monotonous. In the tetrocardin C group, two of the three challenged animals were 9
The animals died on day 1 and day 100, but one animal survived and both drugs were effective. The above results suggest that tetrocardin B and C, like tetrocardin Δ, are effective against BR when administered in small doses, and are also sufficiently effective against Babesia and Theileria in cattle.

Claims (1)

[Scope of Claims] (11) An anti-pyroplasma agent containing at least one of the tetrocardins. (2) An anti-malarial agent containing at least one of the tetrocardins.