FR2509308A1 - MICRONIZED 2- (4-CHLOROPHENYL) -S-TRIAZOLO (5.1-A) ISOQUINOLINE, ITS VETERINARY USE FORMULATION AND METHOD OF USE - Google Patents

Abstract

THE INVENTION RELATES TO 2- (4-CHLOROPHENYL) -S-TRIAZOLO5,1-AISOQUINOLEINE MICRONIZED AND ITS FORMULATION FOR VETERINARY USE. THE MICRONIZED COMPOUND IS MUCH MORE ACTIVE AS A POSTCOITAL ANTI-GESTATION AGENT THAN THE UNMICRONIZED COMPOUND.

Description

In many countries, the accelerated and uncontrolled increase in

  pet population has reached alarming proportions we can admit

  that one of the main causes of this situation is

  in the absence of safe, convenient and cheap methods for controlling the fertility of animals At the present time, only the hormonal compounds used in the veterinary field are used to:

  undesired (estrogen 3) or to presuppose or delay

  During the period of estrus (Prostate and Neglected), however, they are not at risk for immunization and Paina elsewhere, and research into the use of non-hormonal contraceptive agents (eirol and prostaglandins) is not recommended. do not have ibu C auc-iune api licat 11 cn

  practical (J 8 chle, W, Folia "jet L & 4 e 7 GO 5 i 174).

  It has been recently described a number of non-hormonal compounds that are known to be pregnant during pregnancy, at which time there are no animal species after Uterine Tubal Therapy on Uterine Tubes 1 V 7 (1 S-3 A-4,075,341, entitled "Phenyl Compounds <2 Substituted 1-g

  isoquinololein "). These copo are their effectiveness.

  cited during the initial phase of the embryonic

  contraceptive effect is exerted on the uteroplacental ocmplex and results in g nsec and, depending on the animal species, the subsequent estrus or e-xpulsion of the products of conception (G ilan

  et al., Arzneim Forsch, Res., 30, 922, 1980).

  The studies that have been done

  The structure / activity in this class has led to the selection of the compound consisting of 4-amino-5-yl-1,5-azoquinoline and having the following formula:

NOT

  M 1 as one of the most active compounds of the entire class As has been reported in the above-mentioned US patent, this compound exhibits a remarkable activity

  contraceptive when given in multi-dose

  More particularly, US-A-4,075,341 reports the results of experiments on female hamsters by administering this compound, dissolved

  in sesame oil for five consecutive days

  during the peak activity period (of the day

4 to day 8 of pregnancy).

  This result, reported in Table II of this US patent, shows that by giving a daily dose of 0.08 mg / kg, the termination of pregnancy on 50% of the hamsters (ED 50) is obtained. rats by administering the compound dissolved in sesame oil, from day 6 to day 10 of pregnancy, gave an ED 50 of 1 mg / kg / day Despite the very high contraceptive activity demonstrated by these

  tests, the need for multiple administration and over-

  all the need to administer this compound during a

  particular time interval (post-implant phase)

  have rendered this compound unsuitable for

veterinary surgeon.

  It has now been found that by using the particular form, ie the micronized or micropulverized form, the compound 2- (4-chlorophenyl) -5-triazolo [1-isoquinoline] suitable formulations can be prepared for discharge

  prolonged, eliminating the disadvantages mentioned above.

Said formulation, containing

  2- (4-Chlorophenyl) -5-triazolo / 1-ajisoquinoline half

  cronisée, allows the compound to be very effective after a single intramuscular injection, over a longer period of pregnancy Studies on dogs showed that, depending on the dose administered, this period can run of the day mating (or from 10 days before

  mating) on day 15-20 of pregnancy.

  2- (4-Chlorophenyl) -5-triazolo / 5,1-a J-

  micronized isoquinoline is an impor-

  odorless and tasteless, insoluble in water (<1 y / ml) and, at room temperature, moderately soluble in oily vehicles (solubility in sesame oil: 0.5 mg / ml) micronized product has the same analytical characteristics as the non-micronised product, such as for example the same UV spectrum and the same chromatographic Rf, although its

  particles have different dimensions.

  The dimensions of the product particles were measured with the HIAC system. In this system, particle size determination is based on the shadow cast by a particle that is

struck by a ray of light.

  The measuring instrument consists essentially of

  of a sensor which is formed by a light source

  and a photodetector, separated by the counting cell An aqueous suspension of the test powder passes through this cell, the dimensions of which vary according to the range of dimensions to be evaluated. Each particle individually interrupts a part of a

  light beam by generating a signal that is proportional

  on the surface of the shadow of this particle.

  Suitably correlated with the diameter of a standard spherical particle which would give the same light absorption, this electrical signal is filtered and gives the number of particles having a diameter chosen in advance. The instrument can divide the measuring range ( 1 to 300 A) in arbitrarily predetermined size intervals by this method, it is possible to calculate the number of particles for

  each measurement range and put that number in correlation

  the total number of particles in the sample. The values thus obtained are then converted into a single value which represents the

  particle size of the powder.

  The value that is used most commonly

  To represent the particle size of a powder, according to the HIAC evaluation, is the median volume diameter (MDD) which corresponds to the value of the median diameter (50% by volume) of the cumulative volume. The particle size of the

  2- (4-Chlorophenyl) -5-triazolo ,, 1-alisoquinoline micro-

  nized, expressed in DVM, are always less than 15 A.

  2- (4-Chlorophenyl) -5-triazolone is prepared

  Z 5,1-alisoquinoline micronized by subjecting to a mechanical micronization operation, the compound 2- (4-chlorophenyl) -5-triazolol, 1 isoquinoline, obtained by any method described in the technical literature (see for example US Pat. No. 4,075,341 and Italian Patent Applications 26868 A / 79 and 26803 A / 79). This micronization can be carried out using machineries based on various principles, as is well known to those skilled in the art. preferred embodiment of the invention, this micronization is performed in a mill or micronizer fluid energy With this device, the compound to be micronized is trained in

  a circuit by a violent gas flow The colli-

  The particles of the compound against the walls of the circuit as well as the collisions of the particles between them cause the spraying of the particles.

  The apparatus may also be equipped with a recycling device

  The main advantage of the fluid energy mill lies in the fact that the particles of the micronized powder thus obtained have very homogeneous dimensions, that is to say say

  that the particle size range is very narrow.

  2- (4-Chlorophenyl) -5-triazolo, l, -a-

  micronized isoquinoline may be suitably formulated

  in galenic preparations which are adapted to the administration

  parenteral administration, the dimensions of the particles of the initial powder being maintained, and which allow a slow absorption of the active principle, which prolongs its action The formulation of 2- (4-chlorophenyl) s-triazolor 5,1 iia 7 Isoquinoidine micronized - the form of a suspension in a suitable oily vehicle, is therefore another object, specific to the present invention. As a vehicle, it is possible to use oils of vegetable origin. Fat and fatty oils as well as sesame oil, related to cotton oil, and 4-ethyl acetate, are also well known. It is well known to specialists in this field that -imicrobials, -diii so-nt ul Lspour e mp & coher the growth of dir-t years in prepa: ation, and antioxidants, essentitllewment for (exapp Dear the O

oily vehicle of rai-i.

  These suspicions can be estimated at 10%, preferably around 7%.

  active by volume, the opacity of the LMA

  tense of the dosze chosen according to the characters and / or the size of the wool 1 In view of the influence of the invention (dimensions of particles and vehicle) on ato

  contraceptive, ops 2-e 4-ehcrc ohdr-yl) -5-

  triazolorb, 1-azisoquexcjioléin (e to a grinding ilia-nuie and

  an operation m 4 canti d'Aque pirnt'Di re-

  In these cases, the piling of one of the mortars, and one of the mortars, was carried out in one of the mortars and mortars, and this was done in the form of openings which were sufficiently homogeneous. In the second of these methods, the Micronett 1-e M 50 micronizer (GUSEO, Milan, Italy) was operated using a feed pressure of 2 Iar S and a micronization pressure of 7 kV.

  By the methods described above, three

  batches of 2- (4-chlorophenyl) -5-triazolo E 5.1-ajisoquinoline, the particles of which have the following dimensions respectively: 10 A, 39 A and 134 A These three batches of 2- (4-chlorophenyl) -5- triazolo 5,1-a 7 isoquinoline were formulated (as a suspension containing 5% of active ingredient) on the one hand in an oily vehicle (sesame oil containing 0.2% of benzyl alcohol)

  and on the other hand in an aqueous vehicle (physical solution

  siostatic acid containing 2% Tween 8 A. Firstly, the influence of the particle size of the active ingredient and that of the injection vehicle on the embryoletal action of

  compound Sprague-female rats were used for this purpose.

  Dawley, weighing 200-280 g These rats were housed with adult males overnight The following morning,

  the couplings have been highlighted by the pre-

  Sperm Semen in Pap Smear On the same day that sperm was detected (day 1 of pregnancy), rats were administered intramuscularly

  a dose of 10 mg / kg of 2- (4-chlorophenyl) -5-triazolone

  Z ', 1-avisoquinoline The effects of

  on the conceptus by killing animals on day 16 of pregnancy At autopsy, the uteri were isolated

  and determined the number of implantation sites, the name

  number of dead embryos or fetuses and the number of

live fetuses.

  It was considered that the pregnancy was interrupted when the uterine tubes did not contain live fetuses and the presence of implantation scars indicated that the female rats had been fertilized.

  The results obtained on the female rats are summarized

Table I.

TABLE I

  Influence of particle size on the activity of 2- (4-chlorophenyl) -5-triazolo, -i, 1-fluoroquinoline in female rats, following administration of 10 mg / kg on day 1 of pregnancy. (: Dimensions: Number:% Interrup-) (Vehicle: particles: from: tions) (: (DVM) * rates: pregnancy)

(: ::)

(:: :)

(Sesame oil +: 10: 20: 65)

(:: :)

  (0.2% benzyl alcohol: 39: 10: 30)

(:: :)

((0.2 ml / kg): 134: 15: 6.7)

(: :-)

() (t 10 ** Q

(up to:)

  (: :) (Physiological solution :: :) (+: 10: 10: 20) (i :: :) (2% Tween 80 :: :) ((0.2 ml / kg): 39: 6 16.7)

(:::)

(: 134: 11: O)

(: 6 :)

(:: 6 ** O)

(: :)

  * Median volume diameter, expressed in A

  * Animals treated with the vehicle only.

  By analyzing these results, it can be seen that the highest activity occurs when micronized 2- (4-chlorophenyl) -5-triazolo, 1al isoquinoline is administered, suspended in the oily vehicle.

  vehicle, it has been observed that activity decreases progressively

  as the size of the crystals increases.

  On the contrary, the results obtained with the three

  Aqueous vehicles are all negative.

  In view of the above results, we performed

  also experiments on bitches, studying the

  oily suspensions only and by administering the compound at doses four times lower than

  those used in rats, that is to say 2.5 mg / kg.

  This choice has been suggested by the fact that bitches are much more sensitive than rats to the action of these new contraceptives (G Galliani et al Acta Endocrinol.

, Suppl 212,199,1977).

  In this study, a total of 24 adult bitches, of different breeds, weighing from 6 to kg were used. During the estrus phase, the animals were

  mated, then again two days later, with -

  males of proven fertility; the day between the two matings is considered to be the zero day of pregnancy Dogs were administered a single im dose of 2.5 mg / kg during the first six days of gestation and were kept under observation to see if they had ended their pregnancy

or if it was interrupted.

  The results of the experiments carried out on the bitches are reported in Table II:

TABLE II

  Influence of particle size on the activity of 2- (4-chlorophenyl) -5-triazolo-5,1-ajisoquinoline in

  bitches administered 2.5 mg / kg i m.

  suspended in an oily vehicle * during the six

first days of gestation **.

  Dimensions Number of:% Interruptions) particles) (Dv ^)> *** pregnancy bitches)

(10 10: 100

(::)

39 4 50

(:)

134 4 O

(::)

  (vehicle 6 O = to * sesame oil containing 0.2% benzyl alcohol t 0.05 ml / kg)

  ** One injection in the period indicated.

  *** Median volume diameter expressed in A. In agreement with the results obtained for the female rats, the best results (100% of pregnancy interruptions) are also obtained, in dogs with the suspension containing the micronis compound was repeated The above-described experiments used formulations containing Thiophosphonates with dimethylenes (MVD = 14.4 g) and MMVL (k) from 5% results. This year's studies are 5

formulation ontre "i-c l Zci -

  range of 2.5-ng / kg during the 15th of May (from day 0 to day 15), in the case of a break in the survey. 'back E, -more

  e 11 e v e 6 e (5 i g / k q S f icac-

  (93% of rs Vapsti; ï sr between 'durie dy 3vj t the aur

20 of pregnancy.

  uin Zuireouirme 4 J-t ix tion is dolic a u ile * irI '

  unwanted in the case of PZ

  surrendered consists of deducting the inep'-e 11 cz: nzu

  injection and use of the

  the parts of which are less than 15 g and of which they are oily.

Claims (5)

  1- (4-Chlorophenyl) -5-triazolo L / 5,1-a 7-   micronized isoquinoline, the particles of which have a median volume diameter (DVM) size of less than 15 A. 2 A veterinary formulation which contains the compound of claim 1 'as an active ingredient, suspended in an oily vehicle.   Formulation according to claim 2 which   contains 1 to 10 0% (weight / volume) of active ingredient.   Formulation according to claim 3 which   contains about 5% (weight / volume) of active ingredient.   Formulation according to any one of   Claims 2 to 4, characterized in that the   Oily vehicle is sesame oil.   6 Process for interrupting pregnancies   unwanted in pets, which is   to administer to the animal, in a single intramuscular injection, an effective amount of   2- (4-chlorophenyl) -5-triazolo 5,1-isoquinoline   whose particles have dimensions, expressed in DVM, of less than 15 gs and which are suspended in a oily vehicle.    At 8 ': -T M LEM Ol NB nd Batignolle 9 O 9 & W