COMPOSITION FOR ENHANCING CHANCES OF CONCEPTION AND FEMALE SEX RATIO IN BOVINES

Abstract

The present invention includes an economic, synergistic, non-toxic and stable oral composition comprising curcuminoid or its derivative, polyhydric alcohol, amino acids, buffering agent and other pharmaceutically acceptable excipients for enhancing the female sex ratio and improved chances of conception in Bovines. The oral composition is acidic in nature with pH between 4.1-4.6 and in semi-viscous form to enable the availability of the composition in buccal cavity for at least 2-3 minutes. The present invention also provides an economic process for the preparation of said composition.

Claims

1. A synergistic oral composition for enhancing chances of conception and female sex ratio in bovines, wherein the composition comprising curcuminoid or its derivative, polyhydric alcohol, amino acids, or its derivative, buffer agents and other pharmaceutically acceptable excipients.

2. The synergistic oral composition as claimed in claim 1, wherein the composition comprises Demethoxycurcumin in the range of 5-10%, Sorbitol in the range of 10-30%, amino acids in the range of 2-4%, buffering agent in the range of 12-16% and other excipients including water in the range of 48-62%.

3. The synergistic oral combination as claimed in claim 1, wherein the polyhydric alcohol is selected from a group consisting of pentaerythritol, xylitol, adonitol, arabitol, mannitol, sorbitol, dulcitol or other carbohydrate based polyhydric alcohol, preferably the polyhydric alcohol is selected from Sorbitol in the range of 10-30%.

4. The synergistic oral combination as claimed in claim 1, wherein the composition comprises stabilizers selected from a group consisting of glycine, alanine, valine, leucine, isoleucine, phenylalanine, aspartic acid, glutaric acid, asparagine, glutamine, arginine, lysine, cysteine, threonine, tyrosine, praline, histidine, tryptophan, methionine and serine or combination thereof, preferably the amino acid is selected from glutamic acid, aspartic acid or combination thereof, the percentage of 1-2% individually and together 2-4%.

5. The synergistic oral combination as claimed in claim 1, wherein the buffer is selected from citrate buffer or acetate buffer and pH of the composition ranges from 3.5 to 5.5, preferably 4 to 5, most preferably 4.1-4.6.

6. The synergistic oral composition as claimed in claim 1, wherein the excipients are selected from the group comprising binder, stabilizing agent, flavoring agent, sweetening agent, viscosity modifying agent, water or any other compound which is not disturbing the pH of the resulted final composition material.

7. The synergistic oral composition as claimed in claim 1, wherein the composition: i. exhibits high stability at varied temperature range between 6-30° C., ii. is semi-viscous form and retained for 2-3 minutes in the Buccal cavity, iii. mixes with the saliva in 5 to 6 minutes, flow rate of saliva is increased by 10-12 times and it remained higher 2.7 times more when compared to the control group, iv. the pH of the saliva is also dropped to 4.7 from 7.2 and remained at 4.7 for more than 96 hours.

8. The synergistic oral composition as claimed in claim 1 for use in enhancing the conception rate in dairy animals.

9. A process for the preparation of the synergistic oral composition as claimed in claim 1, wherein the said process comprises the following steps of: i. Mixing of Amino acid and curcuminoid or its derivative with water to prepare a mixture, ii. Mixing of both polyhydric alcohol and buffer agent in water to obtain a mixture, iii. Blending of mixtures from step (i) and (ii) together with pharmaceutically acceptable excipients, iv. Evaporating excess water under vacuum to yield the composition in the semi-viscous form.

10. The synergistic oral composition as claimed in claim 1, when orally administering to the bovine once 24 to 48 hours before natural or artificial insemination in a dose of between 0.2-1.0 g/kg body weights of the subjected bovine.

11. A method of increasing the sex ratio in a bovine, said method comprising orally administering to the bovine once the composition as claimed in claim 1 in a dose between 0.2-1.0 g/kg body weights, during 24 to 48 hours before natural or artificial insemination of the subject bovine.

12. The method as claimed in claim 11, where the dose up to 320 grams can be administered to a bovine of 600 Kg.

Description

DETAILED DESCRIPTION OF THE PRESENT INVENTION

[0021] Demand for the dairy products in India and other countries is expected to increase in the coming years driven by more consumers, increase in income and health consciousness. With poor fertility in cows and increasing number of unproductive male cows, it may not be possible to meet the global demand for the dairy products. There is a necessity in developing an alternative method to increase the milk production by enhancing the conception rate and also by altering the sex ratio to especially female offspring in the bovines without causing any change in the genetic profile of the offspring.

[0022] The present invention provides a simple, economic, safe, efficient and synergistic oral composition including curcuminoid or its derivative, polyhydric alcohol, amino acids, or its derivative, buffer agents and other pharmaceutically acceptable excipients to achieve the enhancement in the percentage delivery of female offspring and in the conception rate in bovines.

[0023] The oral composition of the present invention particularly comprises of Demethoxycurcumin in the range of 5-10%, Sorbitol in the range of 10-30%, amino acids in the range of 2-4%, buffering agent in the range of 12-16% and other excipients including water in the range of 48-62%.

[0024] The curcuminoid included in the present composition is selected from a group consisting of curcumin or its derivative, preferably selected from demethoxycurcumin.

[0025] The polyhydric alcohol in the present composition is selected from the group consisting of pentaerythritol, xylitol, adonitol, arabitol, mannitol, sorbitol, dulcitol or other carbohydrate based polyhydric alcohol, preferably the polyhydric alcohol is selected from Sorbitol. The polyhydric alcohol plays a very important role of the carrier in the product delivery system.

[0026] The amino acids used in the present composition are essentially to act as stabilizers and are selected from a group consisting of glycine, alanine, valine, leucine, isoleucine, phenylalanine, aspartic acid, glutaric acid, asparagine, glutamine, arginine, lysine, cysteine, threonine, tyrosine, proline, histidine, tryptophan, methionine and serine or combination thereof, preferably the amino acid is selected from glutamic acid, aspartic acid or combination thereof. The percentage range of amino acid in the composition can be varied between 1-2% individually and together 2-4%.

[0027] The buffering agents of the present composition may be selected from buffer agent, which can provide acidic pH such as citrate buffer or acetate buffer so as to regulate the pH of the composition between 3.5 to 5.5, preferably 4 to 5, most preferably 4.1-4.6.

[0028] The other excipients used in the present invention for making the composition suitable for administration in bovines may be selected from binder, stabilizing agent, flavouring agent, sweetening agent, viscosity modifying agent, water or any other compound which is not disturbing the pH of the resulted final composition material.

[0029] The composition of the present invention exhibits high stability at varied temperature range between 6-25° C. and hence, it requires no special arrangement/instrument like lyophilizer etc for storage. The composition is particularly in semi-viscous form so that it becomes easy for the subjected animal to retain the composition for at least 2-3 minutes in the Buccal cavity for chewing purpose and mixes well with the saliva. By retaining the composition for 2-3 minutes in the buccal cavity, flow rate of saliva is increased by 10-12 times and it remained higher 2.7 times more when compared to the control group. Further, the pH of the saliva is also dropped to 4.7 from 7.2 and remained at 4.7 for more than 96 hours. The increased saliva rate and drop in pH condition assist in creating favourable condition which facilitates the conception possibility in the bovines and particularly towards the birth of female offspring.

[0030] The composition of the present invention: [0031] i. exhibits high stability at varied temperature range between 6-30° C. [0032] ii. is semi-viscous form and retained for 2-3 minutes in the Buccal cavity. [0033] iii. mixes with the saliva in 5 to 6 minutes, flow rate of saliva is increased by 10-12 times and it remained higher 2.7 times more when compared to the control group. [0034] iv. the pH of the saliva is also dropped to 4.7 from 7.2 and remained at 4.7 for more than 96 hours.

[0035] The efficacy of the composition of the present invention is studied in bovines including Heifers and its subsequent parturition. The percentage delivery of female offspring is increased from 45% obtained in the control group to 80% and even in 2.sup.nd or 3.sup.rd parity, Heifers administered with the present composition showed equally good results. The conception rate is also found to have enhanced with the application of the present composition by following the standard protocol of the product administration.

[0036] The composition of the present invention is safe and non-toxic and showed no side effects such as gastrointestinal diseases, uneasiness, fever, skin rashes etc. in the mother animal or in the female offspring produced. More importantly, there is no genetic disorder observed in both mother animal and female offspring produced.

[0037] The present invention also discloses a process for the preparation of the present composition, wherein the said process comprising the steps of: [0038] i. Mixing of Amino acid and curcuminoid or its derivative with water to prepare a mixture. [0039] ii. Mixing of both polyhydric alcohol and buffer agent in water to obtain a mixture. [0040] iii. Blending of mixtures from step (i) and (ii) together with pharmaceutically acceptable excipients. [0041] iv. Evaporating excess water under vacuum to yield the composition in the semi-viscous form.

[0042] The process parameters including temperature and pressure which are room temperature and normal atmospheric pressure.

[0043] The process for the preparation of oral composition of the present invention is carried out in an aseptic environment to avoid any contamination in the prepared composition or to avoid any infection or inflammation in the animal treated with the present composition. The pH of the composition is maintained between 4.1-4.6.

[0044] The present invention also includes various dosage forms of the composition other than the desirable semi-viscous form. For example, the composition material can be in the form of powder, tablet, dispersible tablets or liquid etc. The route for administration of the disclosed composition is basically depends on the subject. In the present invention, subject being the bovine, the composition may be administered via oral administration, particularly through Buccal cavity. Other routes of administration of the present composition may include intravenous, cutaneous, sub-cutaneous, parenteral, vaginal, intra-vaginal, anal routes etc. The composition may be applied either directly or formulated in any desired physical forms.

[0045] The present invention further describes about the dose of administration of the oral composition which may vary according to the body weight of the bovine animal. The prescribed dose for mammal is generally between 0.2-1.0 g/kg body weights of the subjected animal. Accordingly, the amount of present formulation to be given to cows and buffaloes with body weight of 400-600 kg may vary between 250-300 Gms. The application of a very high dosage of composition may lead to lethal effect or serious side effect to the animal.

[0046] Apart from describing suitable dosage form, routes for administration and dosage quantity of the present composition, the present invention further discloses the perfect time for administration of composition, which is crucial for bringing out the better result of the composition i.e. 30 minutes before insemination or at least within one hour after insemination. If the material is administered after this prescribed crucial time, then the composition ingredients may not be available to the animal before ovulation and may not produce the desired result of birth of female offspring.

[0047] In yet another embodiment, the present invention relates to a method of increasing sex ratio in bovine said method comprising orally administering to the bovine once the composition of the present invention, during 24 to 48 hours before natural or artificial insemination in a dose of between 0.2-1.0 g/kg body weights of the subject bovine.

[0048] Accordingly, the amount of present formulation to be given to cows and buffaloes with body weight of 400-600 kg may vary between 250-300 g. It is envisaged that a dose up to 320 gms can be administered to a bovine of 600 Kg.

[0049] The significance of having specified components in the oral composition is as follows: [0050] 1. Presence of polyhydric alcohol preferably Sorbitol increases the flow rate of saliva because of its chewing in the Buccal cavity which helps in reducing pH of the saliva from 7.2 to 4.7 and thus provides favorable conditions for conception and to the birth of female offspring in bovines. [0051] 2. Curcuminoid or its derivative, preferably Demethoxycurcumin acts as an anti-inflammatory agent to prevent any inflammation and infection in the administered animal. It also acts as an immunomodulator and helps animal to have healthy and infection free reproductive organs and zygote formation site without affecting the pH. [0052] 3. Buffer agent helps in regulating and maintaining the pH of the composition between 4.1-4.6. [0053] 4. Presence of amino acid (glutamic acid and aspartic acid) to an extent helps as a stabilizer and also in creating an environment that will create negatively charged ovum to attract positively charged X chromosome bearing sperm and thus producing XX zygote i.e. a female. [0054] 5. Ingredients used in the composition are stable and safe and therefore the composition does not require any costly equipment for storage such as lyophilizer and shows no side-effect.

[0055] The following examples are given as particular embodiments of the present invention and to demonstrate the practice and advantage thereof. It is to be understood that the examples are given by way of illustration and are not intended to limit the specification or the claims that follow in any manner.

EXAMPLE 1

Composition C1-C5 of the Present Invention

[0056] Various illustrative compositions of the present invention are provided at Table 1 below:

TABLE-US-00001 TABLE 1 Composition of the present invention Ingredients C1 C2 C3 C4 C5 Demethoxycurcumin 8 5 7 9 10 Sorbitol 22 15 30 20 10 Aspartic acid 2 1 1 2 1 Glutamic acid 2 1 2 1 1 Buffering Agent 16 16 12 12 16 Total Active Ingredients 50 38 52 44 38 Others & Water 50 62 48 56 62 Total 100 100 100 100 100

EXAMPLE 2

General Procedure for the Preparation of C1-C5 Composition of the Present Invention

[0057] The C1-C5 composition of the present invention is prepared by mixing the amino acids, demethoxycurcumin and water to obtain a first mixture. Then, sorbitol and suitable buffering agent are mixed with water to get a second mixture. Both mixtures are blended together with pharmaceutically acceptable excipients followed by the evaporation of excess water to obtain the formulation in the desired dosage form for oral administration. The pH of the resulting material is maintained between 4.1-4.6 with the use of suitable buffer agents. The process is generally carried out in an aseptic environment.

EXAMPLE 3

Preparation of Composition of the Present Invention

[0058] Aspartic acid (2.0 g), glutamic acid (2.0 g) and Demethoxycurcumin (8.0 g) are mixed well with water (20.0 g) to get a first mixture. Then, Sorbitol (22.0 g), citrate buffer (16.0 g) and water (20.0 g) are mixed together to prepare a second mixture. Both mixtures are blended together with maltose excipients (5.0 g) and glycerin (5.0 g), concentrated under vacuum to remove excess water to obtain the composition in a semi-viscous form.

EXAMPLE 4

Preparation of Composition of the Present Invention

[0059] Aspartic acid (1.0 g), glutamic acid (2.0 g) and Demothoxycurcumin (7.0 g) are mixed well with water (20.0 g) to get a first mixture. Then, Sorbitol (30.0 g), acetate buffer agent (12.0 g) and water (20.0 g) are mixed together to prepare a second mixture. Both mixtures are blended together with maltose excipient (5.0 g) and glycerin (3.0 g), concentrated under vacuum to remove excess water to obtain the composition in a semi-viscous form.

EXAMPLE 5

Effect of the Components of the Present Invention

[0060] Several compositions were made by varying the components of the present invention and the same are elaborated in Table 2 below:

TABLE-US-00002 TABLE 2 Testing of importance of the composition of the present invention Ingredients F1 (C1) F2 F3 F4 F5 Demethoxycurcumin 8 0 7 9 10 Sorbitol 22 20 0 20 10 Aspartic acid 2 1 1 0 1 Glutamic acid 2 1 2 0 1 Buffering Agent 16 16 12 15 0 Total Active Ingredients 50 38 82 44 54 Others & Water 50 62 48 56 62 Total 100 100 100 100 100

[0061] The F1 is per the composition C1 of the present invention. The compositions F2 to F5, miss one of the essential components of the composition. The above table clearly shows the interaction between the essential ingredients to provide the surprising efficacy of the components of the composition and shows the deleterious of effect of altering the components of the composition.

[0062] The compositions F1 to F5 were tested by Veterinarians in the field. It was found that the composition F1 was the only active composition. The compositions F2 to F5 were not formed into a stable mixture or were not active. F1 also gave the best results as the availability all the ingredients ensured better and consistent results. The absence of Demethoxycurcumin in F2 resulted in the inconsistent uterine pH. The absence of Sorbitol in F3 had lesser stability and was not available in the buccal cavity for longer duration. The absence of the mentioned amino acids in F4 resulted in lesser motility of the sperms in the uterus. The total absence of the Buffering agents in F5 did not provide the desired pH.

[0063] It has been observed that when there is change in the physicochemical parameters, there is a change in efficacy and only the composition Cl of the present invention. From the above it can be clearly seen that unless all components of the composition are present the desired results of altering the sex ratio cannot be achieved. In other words, the composition is synergistic showing unexpected efficacy by the interaction of its ingredients. This composition and its efficacy are arrived after much human efforts and several clinical trials.

EXAMPLE 6

Activity of the Composition of the Present Invention

[0064] The efficacy of the present composition in enhancing conception ratio and female sex ratio in diary animals is tested on initial trial basis in bovine animals. During the trial testing, the composition is administered as per the standard protocol and in the presence of supervision of qualified Veterinarians, Breeders and Applicators. The results are tabulated in Table 3.

TABLE-US-00003 TABLE 3 Effect of the composition of the present invention Table 3A: Effect in Heifers Heifers With new Composition Controlled AI (Artificial 28 25 insemination) Pregnancy 20 11 Conception Ratio 71.43% 44.00% Female 16 80.00% 5 45.45% Male 4 20.00% 6 54.55%

[0065] The birth of female offspring in case of heifers administered with the new composition is up-to 80% compared to 45.45% in the controlled group. The conception ratio of heifers subjected to present composition is also improved to 71.43% from 44%.

TABLE-US-00004 TABLE 3B Effect in Animals in 2-3 parity 2.sup.nd -3.sup.rd Parity With new Composition Controlled AI 35 40 Pregnancy 23 19 Conception Ratio 65.71% 47.50% Female 18 78.26% 9 47.37% Male 5 21.74% 10 52.63%

[0066] The birth of female offspring in case of 2.sup.nd and 3.sup.rd Parity administered with the new composition is up-to 78.26% little lesser in comparison to Heifers and compared to controlled group it is much higher at 47.37%. The conception ratio of heifers subjected to present composition is also improved to 65.71% from 47.50%.

TABLE-US-00005 TABLE 3C Effect in Heifers & Animals in 2nd-3rd parity Total AI With new Composition Controlled AI 63 65 Pregnancy 43 30 Conception Ratio 68.25% 46.15% Female 34 79.07% 14 46.67% Male 9 20.93% 16 53.33%

[0067] Table 3B and 3C clearly indicate that the composition of the present invention is quite effective in improving the chances of producing female off-springs even in 2.sup.nd or 3.sup.rd parity animals including Heifer animals in 2.sup.nd or 3.sup.rd parity. The percentage of delivery of female calves in Heifers and animals in 2.sup.nd or 3.sup.rd parity cumulatively is around 79.07% compared to 46.67% via controlled method. The conception rate is also increased up to 68.25% with the application of the composition of the present invention compared to 46.15% achieved via controlled method.

[0068] The toxicity effect of the present composition is also studied in bovines. The composition is found to be safe and non-toxic and showed no side-effects such as gastrointestinal disorder, genetic disorder, increase in body temperature and uneasiness etc. in both subjected animal and its offspring.

Advantages Of The Present Composition

[0069] Composition is highly palatable product with soothing flavor.

[0070] Composition shows better stability at a temperature range between 6-25 ° C.

[0071] Composition can be easily metabolized and thus eliminates chance of any genetic disorder.

[0072] Composition may be administered just on the onset of the heat of the animal.

[0073] Composition provides balanced pH at the zygote formation site and stability for enhancing female ratio in Bovines.

[0074] It provides no side effects on mammals after the application.

TABLE-US-00006 TABLE 4 Effect of the individual Components in the composition of the present invention Ingredient Activity Sorbitol 10% Aspartic acid  3% Glutamic acid  3% Acetic acid 23% Acetate buffer 21% Demethoxycurcumin 5-7% Maltose Nil Composition of the present invention 83%

[0075] From the above table it is very clear that the addition of the active ingredients like Aspartic acid and Glutamic acid have some effect on the performance of the product by supplementing the energy to the sperms. The presence of the derivatives of Curcumin Demethoxycurcumin ensures better health to the animal and also provides immune-modulatory benefits to the animal. The presence of Demethoxycurcumin also provides better motility to the sperms. The entire composition helps to maintain the pH of the Reproductive organs to the acidic as the post ovulation pH of the Ovum is more likely alkaline and the uterine pH is expected to be 6.7 to 7.2. The present composition ensures to have a negatively charged environment thus enabling the higher chances of having XX zygote formation.

EXAMPLE 7

Effect of the Composition of the Present Invention in Female Ratio in Cows

[0076] The composition of the present invention of Polyhydric Alcohols, Amino acids, Maltose, Demethoxycurcumin and buffers of acetates as illustrated at tables 5 and 6 below was tested for its efficacy in cows (Table 5) and in Buffaloes (Table 6) after obtaining necessary ethical clearances. The composition was administered at a dose of 320 grams per animal single dose of the composition at the on-set of heat symptoms, i.e. at the onset of oestrous cycle.

TABLE-US-00007 TABLE 5 Effect of the composition of the present invention on cows Number of animals Number with Sex of new- Sr. of animals Confirmed Calving born calf No. Group No. in group pregnant occurred Female Male 1 Group-1 40 25   23   19   4   Percentage 62.5  92   83   17   2 Group-2 40 26   25   20   5   Percentage 65   96   80   20   3 Group-3 40 21   18   16   2   Percentage 52.5  85.7  89   11   4 Group-4 40 18   15   14   1   Percentage 45   83   93   7   5 Group-5 40 21   19   15   4   Percentage 53   90   79   21   6 Group-6 40 32   28   23   5   Percentage 80   87.5  82   18   7 Controlled 40 16   15   8   7   Group Percentage 40   93.75 53.33 46.67

TABLE-US-00008 TABLE 6 Effect of the composition of the present invention on Buffaloes number of Number animals of confirmed Calving sex of new-born calf Group No. animals Pregnancy occurred Female Male Group-1 40 20 19 17 2 Percentage 50 95 89 11 Group-2 40 23 23 19 4 Percentage 58 100 83 17 Group-3 40 15 15 13 2 Percentage 38 100 87 13 Group-4 40 21 19 17 2 Percentage 53 90 89 11 Group-5 40 15 14 13 1 Percentage 38 93 93 7 Group-6 40 19 18 16 2 Percentage 48 95 89 11 Controlled Group 40 15 14 7 7 Percentage 37.5 93.33 50 50

[0077] From Tables 5 and 6 as above, it can be seen that the Composition in both Cows and Buffaloes demonstrates enhanced conception and an increased female ratio of 80% on various parameters with the ratio of the female calves as high as 93% in comparison to the controlled group where the conception was as low as 37.5% in Buffaloes and 40% in cows and the female ratio in Buffaloes was 50% and in cows it was 53.3%.

EXAMPLE 7

Comparison of the Composition of the Present Invention with Prior Art Composition

[0078] The composition of the present invention is compared with a prior art composition and other compositions such as in Aulprofem and Ratio, and the results are provided as here below at Table 7.

TABLE-US-00009 TABLE 7 Showing the comparison of the composition of the present invention with prior art composition P1 P2 (Aulprofem) (Ratio) Present composition (C1/F1) Acetic Acid 10% Sorbitol   64% Demethoxycurcumin  8% Sodium Acetate  6% Aspartic acid  0.5% Sorbitol  22% Glutamic acid  1.5% Aspartic acid  2% Acetate buffer  4.0% Glutamic acid  2% Maltose   15% Buffering Agent  16% Others & Water   15% Total Active Ingredients  50% Total  100% Others & Water  50% Total 100%

The Composition of the Above-Mentioned Invention is Better Than the Two Previous Ones on Various Parameters

[0079] The composition is more stable and consistent due to stability of pH.

[0080] The conception also increased considerably using the composition of the current invention.

[0081] The presence of Curcuminoid or its derivatives ensures the healthy uterine and other reproductive organs as its takes care of any possibility of the infection at the zygote formation site and also acts a strong immune-modulator.

[0082] The overall female births also improved with the current invention.

[0083] Overall results with the current composition of the new invention were better than the previous two.

[0084] From the above and based on clinical trials, it can be seen that the composition of the present invention has enhanced efficacy than prior art compositions and provides better female sex ration in comparison to other compositions of prior art known for such purposes.