SBIR-STTR Award

As their first application for veterinary health, the two PI’s have already remarkably employed their novel, patented superagonist and neoglycosylation technology to develop the first recombinant, much more potent and long-acting bovine FSH antagonists for improved cattle reproduction and a potential $30-40 Million annual market. This work was supported by a previous phase 1 SBIR from FDA-CVM which proved vital in obtaining required data for an FDA NADA and ultimately for a large licensing & commercialization agreement with Zoetis, the world’s leading veterinary health company. The current proposal of the first recombinant, much more potent and long acting equine chorionic gonadotropin (eCG) agonists for improved and more ethical reproduction in pigs and cattle is estimated both by Trophogen and potential partnering veterinary pharma companies to be an even larger annual worldwide market of $50-100 Million, a potential veterinary “blockbuster”! In view of the PI’s remarkable success in developing and commercializing its first veterinary product with support of a phase 1 SBIR from FDA and validation of its technology and the potential market value of its reproductive products by world leader Zoetis, we hope that the current submission will receive expedited approval. The goal of this Phase 1 project is to develop and evaluate new recombinant eCG analogs as replacement for pregnant mare’s serum gonadotropin (PMSG) and P.G. 600® (PMSG+hCG). The use of PMSG obtained by inhumane bleeding of pregnant mares between days 40 and 130 of gestation is highly controversial and an inhumane business of PMSG production raises increasing concerns of the public and animals rights group. There are many limitations of current PMSG and related products including presence of horse serum contaminants in PMSG preparations, potential infection with equine viruses and prions, as well as immunological responses to highly heterogenous preparations produced in heterologous species (horse or human). There are no approved recombinant veterinary eCG products and the PIs were highly encouraged by increasing interest from major veterinary companies in their totally novel more potent and longer-acting recombinant gonadotropins, including new eCG analogs. In this Phase 1 we plan to: Aim1A Develop novel high affinity recombinant eCG superagonist analogs by minimal 4 arginine site-directed mutagenesis to produce a more potent and efficacious dual FSH + LH/CG activity molecule; Aim1B For selected eCG superagonists also engineer a novel minimal length amino acid insert containing one or two complex carbohydrate neoglycosylation sites to further increase half-life and produce analogs without reducing increased superagonist potency/efficacy; Aim 2 Optimize production and purification of these novel analogs using stable highly expressing Chinese hamster ovary (CHO) cell lines and low cost production methods in roller bottles; Aim 3 Select the best 2-4 analog candidates and characterize them in vitro by previously validated eCG analog ELISA immunoassay, SDS-PAGE electrophoresis, isoelectric focusing (IEF) gel analysis and robust in vitro FSH and LH/CG bioassays; Aim 4 Select the final eCG analog using the most important parameter determining in vivo performance: pharmacokinetic (PK) studies in mice; Aim 5 Establish CHO research cell bank (RCB) providing high expression of the lead, optimally neoglycosylated eCG analog sufficient for more expensive bioreactor production and all extensive rodent, pig and cattle studies in Phase 2.

Public Health Relevance Statement:
Pregnant mare’s serum gonadotropin (PMSG=eCG) obtained from “horse blood farms” is the main ingredient of several veterinary products that are artificially inducing heat in weaned sows to achieve a faster and more controlled reproduction. There are many limitations, ethical concerns and risks associated with PMSG, derived from serum of pregnant mares, including possible contamination by a protein infectious agent which causes a 100% fatal brain disease in humans after eating infected meat. The design and development of the first synthetic DNA-derived, highly pure, much more potent and long acting dual-activity eCG analog, which cannot contain any infectious agent, offers not only prospects of largely improved, safe and more ethical reproduction in pigs and cattle, but also a unique potential for veterinary reproduction “blockbuster” with multiple applications in different species!

Project Terms:
absorption; Affinity; Agonist; Agreement; Amino Acids; analog; Animal Rights; Animals; Area; Arginine; Biological Assay; Bioreactors; Blood; Brain Diseases; Businesses; Carbohydrates; Cattle; cell bank; Cell Line; Charge; Chinese Hamster; Chinese Hamster Ovary Cell; Chorionic Gonadotropin; commercial application; commercialization; Complex; cost; Data; design; Development; Diagnosis; Drug Kinetics; Eating; Electrophoresis; Electrostatics; Engineering; Enzyme-Linked Immunosorbent Assay; Epitopes; Equus caballus; Ethics; Family suidae; Farming environment; Follicle Stimulating Hormone Receptor; Food Safety; Food Supply; Gel; Generations; Glycoproteins; Goals; Half-Life; Health; Hemorrhage; hormone analog; Hormones; Human; Human Chorionic Gonadotropin; Human Follicle Stimulating Hormone; Immune response; Immunoassay; immunogenic; improved; In Vitro; in vivo; Infection; Infectious Agent; Injection of therapeutic agent; interest; Isoelectric Focusing; Lead; Legal patent; Length; LH Receptors; Licensing; Malignant neoplasm of thyroid; Masks; Meat; Methods; Mus; Mutagenesis; novel; Ovary; Performance; Peripheral; Phase; PMSG (Gonadotropins); Pregnancy; pregnant; Preparation; Prions; Production; Proteins; receptor; Recombinant Gonadotropin; Recombinants; Reproduction; reproductive; Research; Risk; Rodent; Roller Bottle; Serum; Site; Site-Directed Mutagenesis; Small Business Innovation Research Grant; Structure; success; Superovulation; synthetic construct; Technology; United States National Institutes of Health; Validation; Virus; Weaning; Work

The current proposal of the PI’s second veterinary product, the first more potent and long acting recombinant equine chorionic gonadotropin (reCG) agonist for improved and, for the first time, totally ethical reproduction in pigs and cattle by acceleration to the times of estrus is estimated to be $ 200 M + annual market, a potential veterinary product “blockbuster”! The currently used animal product, pregnant mare serum gonadotropin (PMSG) obtained by increasingly recognized inhumane bleeding of noble pregnant mares is highly controversial, has raised increasing concerns among animal rights activists, regulatory and political bodies which have already or soon plan to ban the sale of PMSG. Our previous Phase 1 eCG SBIR achieved a remarkable impact score of 25 and was funded by NICHD in its first submission. The most remarkable aspects of this current Phase 2 SBIR submission is that, first, we provide compelling data that we have not only achieved but have greatly exceeded all aims of our Phase 1 eCG analog SBIR proposal. Secondly and remarkably, we have already established an extraordinarily advantageous commercialization agreement in Phase 1 with Merck Animal Health, the manufacturer of the leading current PMSG product which vitally wishes to replace its natural eCG with this greatly improved recombinant analog. Our aims are: YEAR 1 Aim 1: Improve expression, glycosylation and a major commercial scale up of its bioreactor production and purification methods: A. State of the art permeable beads with adherent CHO cells in suspension; B. Larger fibrous bed bioreactor as developed in Phase 1; C. Possible adaptation of adherent CHO cells to direct growth in suspension culture; Aim 2: Extensive PK studies in rodents to model optimized dose ranging amount and injection number of in vivo bioassays. YEAR 2 Aim 1: Development of CHO Cell Master Cell Bank under GMP conditions of either adherent or suspension cells suitable for transfer to a Merck Animal Health Commercial Bioreactor Facility; Aim 2: Testing of multiple commercial uses of the eCG in both pigs and cows in parallel with Merck Animal Health under GLP conditions suitable to submit an INAD to FDA-CVM and begin regulatory discussions for Fast Track Approval for human and animal safety and ethical treatment of horses: A. Synchronization and acceleration to estrus in gilts and sows; B. Synchronization and acceleration to estrus in heifers and cows; C. Superovulation of cattle; Aim 3: Testing for presumed immunogenicity of the human CG component within Merck’s PG 600: A. Serum immunogenicity studies to hCG and eCG on single and repeated use; B. Possible attenuation of clinical response in repeated use. In view of the PI’s remarkable success in commercializing its very first veterinary bovine FSH analog product with world leader animal pharma Zoetis and now, secondly, for its eCG analog by number 3 animal pharma, Merck Animal Health, we believe this Phase 2 proposal strongly merits expedited approval.

Public Health Relevance Statement:
The current proposal of the PI’s first recombinant, much more potent and long-acting equine (horse) chorionic gonadotropin (eCG) agonist for improved and, for the first time, totally ethical reproduction in pigs and cattle is estimated to be a very large $ 200 M + annual market, a potential veterinary product “blockbuster”! The goal of this project is to develop and evaluate new recombinant equine chorionic gonadotropin (eCG) analogs as replacement for pregnant mare’s serum gonadotropin (PMSG). The use of PMSG obtained by increasingly recognized inhumane bleeding of noble pregnant mares is highly controversial has raised increasing concerns among animal rights groups, regulatory and political bodies which have already or soon plan to ban the sale of PMSG throughout the world.

Project Terms:
Acceleration; Achievement; Affinity; Agonist; Agreement; analog; Animal Rights; animal safety; Animals; attenuation; Beds; Bioreactors; Cattle; cell bank; Cells; Chinese Hamster Ovary Cell; Chorionic Gonadotropin; Clinical; commercial application; commercialization; Country; Data; Development; Diagnosis; Dose; egg; Equus caballus; Estrus; Ethics; experience; Family suidae; Funding; Generations; Glycoproteins; glycosylation; Goals; Growth; Health; Hemorrhage; hormone analog; Hormones; Human; Human Chorionic Gonadotropin; Human Follicle Stimulating Hormone; immunogenicity; improved; in vivo Bioassay; Infertility; Injections; Licensing; Livestock; Malignant neoplasm of thyroid; Manufacturer Name; Methods; Modeling; National Institute of Child Health and Human Development; novel; Permeability; Phase; PMSG (Gonadotropins); Politics; pregnant; Production; Recombinants; Recording of previous events; Reproduction; Research; response; Risk; Rodent; Sales; scale up; Serum; Small Business Innovation Research Grant; Study Section; success; Superovulation; Suspension Culture; Suspensions; Testing; Time; United States National Institutes of Health; Work