Injectable contraceptive and method

Claim 1

A method of causing a delay or bypass of estrus in a female canine having a known reproductive history consisting of either at least one previous estrus cycle, a previously successful pregnancy or both which comprises parenterally administering to said canine, during late anestrus to mid-proestrus, at least 15,000 picogram equivalent antibody units per pound of body weight of a contraceptive antiserum that has been prepared by a process of: (a) preparing an antigen which is capable of stimulating the immunological system of a female canine to form antibodies against estradiol-17 beta by the esterification of the hydroxyl groups of estradiol-17 beta with succinic anhydride to form the estradiol-17 beta hemisuccinate and coupling said hemisuccinate to the amino groups of a protein selected from the group consisting of keyhole limpet hemocyanin, bovine serum albumin and human serum albumin in an amount such that to each molecule of protein there is attached 500-5000 molecules of the estradiol-17 beta. (b) initially parenterally administrating to a female canine donor at intramuscular sites and multi-intradermal sites 1 mg of said antigen emulsified in 2 ml of an adjuvant therefor, parenterally administering additional doses of said emulsified antigen at periodic intervals to cause the production of a desired level of antibodies to said antigen by the immunological system of said donor; (c) collecting blood from said female canine donor in a manner to prevent coagulation of said collected blood, recovering the plasma of said blood by centrifugation, defibrinating said plasma to obtain antiserum containing said antibodies; and (d) diluting said antiserum in a buffered solution of a preservative therefor to a desired concentration.

Claim 2

A method of causing a delay or bypass of estrus in a female canine having a known reproductive history consisting of either at least one previous estrus cycle, a previously successful pregnancy or both which comprises parenterally administering to said canine, during late anestrus to mid-proestrus, at least 15,000 picogram equivalent antibody units per pound of body weight of a contraceptive antiserum that has been prepared by a process of: (a) preparing an antigen which is capable of stimulating the immunological system of a female canine to form antibodies against estradiol-17 beta by the esterification of the hydroxyl groups of estradiol-17 beta with succinic anhydride to form the estradiol-17 beta hemisuccinate and coupling said hemisuccinate to the amino groups of a protein selected from the group consisting of keyhole limpet hemocyanin, bovine serum albumin and human serum albumin in an amount such that to each molecule of protein there is attached 500-5000 molecules of the estradiol-17 beta. (b) initially parenterally administrating to a female canine donor at intramuscular sites and multi-intradermal sites 1 mg of said antigen emulsified in 2 ml of an adjuvant therefor, parenterally administering additional doses of said emulsified antigen at periodic intervals to cause the production of a desired level of antibodies to said antigen by the immunological system of said donor; (c) collecting blood from said female canine donor in a manner to prevent coagulation of said collected blood, recovering the plasma of said blood by centrifugation, defibrinating said plasma to obtain antiserum containing said antibodies; and (d) diluting said antiserum in a buffered solution of a preservative therefor to a desired concentration.

Claim 9

The method of causing a delay or bypass of estrus in a female canine which comprises parenterally administering to a female canine with a known reproductive history consisting of either one previous estrous cycle, a previously successful pregnancy or both, an effective amount in the order of 15-80,000 picogram equivalent antibody units per pound of body weight during late anestrus to mid-proestrus of the reproductive cycle of a contraceptive antiserum which has been prepared by a process which comprises the steps of: (a) preparing an antigen which is capable of stimulating the immunological system of a female canine to form antibodies against estradiol-17 beta by the esterification of the hydroxyl groups of estradiol-17 beta with succinic anhydride to form the estradiol-17 beta-hemisuccinate and coupling said hemisuccinate to the amino groups of keyhole limpet hemocyanin via the carbodiimide reaction in amounts such that to each hemocyanin molecule 1600-2200 of the hemisuccinate molecules are attached; (b) initially injecting parenterally a female canine donor with 1 mg of antigen, emulsified in 2 ml. of an adjuvant, into multi-intradermal sites and intramuscular sites, followed by secondary injections of said emulsified antigen at periodic intervals for periods of time such that the immunological system of said female canine donor can produce antibodies to said antigen at the desired level; (c) collecting blood from said female canine donor in a vessel containing an anticoagulant, recovering the plasma of said blood by centrifugation; (d) converting said plasma to an injectable antiserum by shaking with calcium chloride to form fibrin, removing fibrin by filtration, diluting said antiserum with a buffered preservative solution to a concentration of 1:10,000 and sterile filtering said antiserum.

Claim 10

The method of causing a delay or bypass of estrus in a female canine which comprises parenterally administering to a female canine with a known reproductive history consisting of either one previous estrous cycle, a previously successful pregnancy or both, an effective amount in the order of 15-80,000 picogram equivalent antibody units per pound of body weight during late anestrus to mid-proestrus of the reproductive cycle of a contraceptive antiserum which has been prepared by a process which comprises the steps of: (a) preparing an antigen which is capable of stimulating the immunological system of a female canine to form antibodies against estradiol-17 beta by the esterification of the hydroxyl groups of estradiol-17 beta with succinic anhydride to form the estradiol-17 beta-hemisuccinate and coupling said hemisuccinate to the amino groups of keyhole limpet hemocyanin via the carbodiimide reaction in amounts such that to each hemocyanin molecule 1600-2200 of the hemisuccinate molecules are attached; (b) initially injecting parenterally a female canine donor with 1 mg of antigen, emulsified in 2 ml. of an adjuvant, into multi-intradermal sites and intramuscular sites, followed by secondary injections of said emulsified antigen at periodic intervals for periods of time such that the immunological system of said female canine donor can produce antibodies to said antigen at the desired level; (c) collecting blood from said female canine donor in a vessel containing an anticoagulant, recovering the plasma of said blood by centrifugation; (d) converting said plasma to an injectable antiserum by shaking with calcium chloride to form fibrin, removing fibrin by filtration, diluting said antiserum with a buffered preservative solution to a concentration of 1:10,000 and sterile filtering said antiserum.