SBIR-STTR Award

Malaria is responsible for several hundred million cases and over one million deaths per year. A highly effective vaccine that prevents blood stage infection and thereby prevents all disease and transmission would be an ideal tool for eliminating Plasmodium falciparum (Pf), the cause of 99% of malaria mortality. Attenuated Pf sporozoites (PfSPZ) administered by the bite of infected mosquitoes, lead to protection of >90% of human volunteers against experimental Pf challenge that lasts at least 10 months. Sanaria's goal is to develop and commercialize an attenuated PfSPZ vaccine that prevents Pf blood stage infection in >90% of recipients - a vaccine with these characteristics could be used to eliminate Pf from the world. This vaccine has the potential for >$1 billion annual revenues. Sanaria has successfully established robust, reproducible, and consistent manufacture and release of clinical lots of its PfSPZ Vaccine, received FDA approval (IND) for clinical trials, and initiated a Phase 1 clinical trial to assess safety, immunogenicity, and protective efficacy of the PfSPZ Vaccine in May 2009. Additional proof of concept studies in African adults, young children, and infants are planned and will be followed by Phase 2 studies, and then pivotal Phase 3 studies to support licensure. Sanaria's manufacturing process utilizes a master cell bank of cryopreserved P. falciparum-infected erythrocytes as a source of parasites, and Anopheles stephensi mosquitoes to raise the PfSPZ. However, because there is no method for cryopreserving any stage of the Anopheles mosquito life cycle, the mosquitoes used for producing the PfSPZ Vaccine come from a mosquito colony that is continuously maintained. Cryopreservation of the eggs of Anopheles spp. would enable Sanaria to cryobank different strains of An. stephensi, reduce costs associated with strain maintenance, produce a uniform seed stock and provide security against strain loss or genetic drift. Cryobanking of transgenic An. stephensi and other Anopheles spp. would also provide direct benefit to those aiming to use genetically altered or sterile mosquitoes for transmission control. In pilot studies, we have shown that An. stephensi can be cryopreserved in the vapor phase of liquid nitrogen using a novel cryoprotectant additive (CPA) mixture, with successful hatching and larval development following thawing. An invention disclosure describing the details that led to the production of the first successful Anopheles egg cryopreservation has been filed. The hatch rate to date has been 10% compared to non-cryopreserved controls from the same egg batch which is clear indication of proof of concept. This proposal will define and optimize the successful cryopreservation of An. stephensi by a pragmatic and systematic testing of embryos at different stages of development and of cryopreservation methods and additives. Successful outcome will be measured as the development from cryopreserved eggs of An. stephensi that retain full susceptibility to infection with PfSPZ in the salivary glands. Other strains of An. stephensi held by Sanaria, including transgenic strains, will be similarly cryopreserved.

Public Health Relevance:
Malaria causes 500 million clinical cases and 1-3 million deaths annually, is responsible for >1% loss of GDP in Africa annually and is a serious concern for travelers and military personnel. Sanaria's goal is to develop and commercialize a >90% protective malaria vaccine for three primary markets with a potential for >$1 billion annual revenues. Success in this project will represent a major breakthrough in the fields of vector and malaria biology, will ensure the ability to conserve the genetic stock of the mosquitoes used by Sanaria's in its vaccine manufacturing process, will move Sanaria towards ensuring consistency of mosquito colonies across different manufacturing sites and will provide a secure cryobank of material from which mosquito production can be initiated periodically and which can be followed in a well-defined stability program.

Thesaurus Terms:
"0-11 Years Old; 21+ Years Old; Adult; Africa; African; Anopheles; Anopheles Genus; Armed Forces Personnel; Attenuated; Biology; Biotechnology; Bite; Blood; Blood Erythrocyte; Blood Normocyte; Cessation Of Life; Characteristics; Child; Child Youth; Children (0-21); Clinical; Clinical Trials; Clinical Trials, Phase I; Clinical Trials, Unspecified; Cryofixation; Cryopreservation; Culicidae; Death; Development; Disclosure; Early-Stage Clinical Trials; Embryo; Embryonic; Ensure; Erythrocytes; Erythrocytic; Fecundability; Fecundity; Fertility; Genetic; Genetic Differentiation; Genetic Divergence; Genetic Drift; Goals; Head And Neck, Salivary Glands; Human Volunteers; Human, Adult; Human, Child; Infant; Infection; Information Disclosure; Institutes; Lead; Length Of Life; Licensure; Life Cycle; Life Cycle Stages; Liquid Substance; Longevity; Maintenance; Maintenances; Malaria; Malaria Vaccines; Malarial Vaccines; Marketing; Marrow Erythrocyte; Maryland; Measures; Methods; Military; Military Personnel; Mortality; Mortality Vital Statistics; Mosquitoes; N Element; N2 Element; Nitrogen; Outcome; Paludism; Parasites; Pb Element; Phase; Phase 1 Clinical Trials; Phase I Clinical Trials; Phase I Study; Pilot Projects; Plant Embryos; Plasmodium Infections; Plasmodium Falciparum; Predisposition; Preparation; Procedures; Process; Production; Programs (Pt); Programs [publication Type]; Protocol; Protocols Documentation; Quality Control; Red Blood Cells; Red Cell; Red Blood Corpuscule; Red Cell Of Marrow; Reticuloendothelial System, Blood; Reticuloendothelial System, Erythrocytes; Sbir; Sbirs (R43/44); Safety; Salivary Glands; Secure; Security; Seeds; Site; Small Business Innovation Research; Small Business Innovation Research Grant; Source; Sporozoite Vaccine; Sporozoites; Staging; Sterility; Susceptibility; Technology; Testing; Transgenic Organisms; Transmission; Universities; Vaccines; Zygotes, Plant; Adult Human (21+); Adult Youth; Blood Corpuscles; Cell Bank; Children; Clinical Investigation; Clinical Lot; Cold Preservation; Cold Storage; Communicable Disease Transmission; Cost; Disease Transmission; Egg; Fluid; Hatching; Heavy Metal Pb; Heavy Metal Lead; Immunogenicity; Infectious Disease Transmission; Life Course; Life Span; Lifespan; Liquid; Manufacturing Process; Novel; Phase 1 Study; Phase 1 Trial; Phase 2 Study; Phase 3 Study; Phase I Trial; Pilot Study; Prevent; Preventing; Programs; Protective Efficacy; Protocol, Phase I; Public Health Relevance; Scale Up; Seed; Sterile; Success; Tool; Transgenic; Transmission Process; Vapor; Vector; Young Adult; Youngster"

The ideal, most effective, tool for eliminating Plasmodium falciparum (Pf), the causative agent of 99% of all malaria deaths, is a highly effective vaccine that prevents blood stage infection and thereby prevents both disease and transmission. Sanaria's goal is to develop and commercialize a Pf sporozoite (SPZ) vaccine that prevents Pf blood stage infection in >90% of recipients. The Sanaria(R) PfSPZ Vaccine is composed of attenuated, purified, aseptic, cryopreserved PfSPZ. The platform technology developed to manufacture the vaccine has also facilitated the manufacture of PfSPZ for challenge infections to test vaccines and drugs (PfSPZ Challenge) and for vaccination by challenge under chemoprophylaxis (PfSPZ-CVac). These comprise a portfolio of products that are on an aggressive timeline to commercialization. Manufacture of all these PfSPZ products is dependent on Anopheles stephensi mosquitoes that can survive aseptic rearing from eggs to adults and produce high numbers (>105) of PfSPZ in their salivary glands. A vulnerable aspect of Sanaria's manufacturing process is its reliance on mosquitoes that can only be maintained in colonies; such colonies are susceptible to crashes and to genetic drift over time, thereby being at risk of permanently losing the desired genotype and phenotype. This is because despite decades of effort, until our breakthrough achievement in Phase I, live Anopheles could not be preserved in any way. In Phase I we demonstrated new enabling technology, the capability to cryopreserve eggs of A. stephensi (SDA500-9800 strain). In the GMP context, being able to cryopreserve large lots of eggs will allow Sanaria to inaugurate controlled manufacturing by use of a Master Mosquito Bank, thereby enabling us to return repeatedly to a repository stock generated from the same defined, characterized material with the desired genetic background. With careful management, such a stock can last for many tens of years. In this Phase II project we will fully optimize the cryopreservation method (Specific Aim 1), and use this method in scaled-up production, initially to ensure security of the colony for manufacturing purposes (Specific Aim 2) and subsequently for producing the larger Master Mosquito Egg Bank (Specific Aim 3) from which all mosquitoes used in manufacturing will be sourced. Details of the characterized Master Mosquito Egg Bank will be submitted to the FDA as a Biologics Master File. We will also systematically apply the egg cryopreservation technology to support development of improved strains of A. stephensi and to create banks of eggs of such strains when developed (Specific Aim 4). Success in this project will have a critically important impact on Sanaria's commercial success.

Thesaurus Terms:
Achievement;Adult;Africa;African;Anopheles Genus;Antimalarials;Attenuated;Back;Base;Biological Preservation;Bite;Blood;Cessation Of Life;Chemoprophylaxis;Chloroquine;Clinical;Clinical Trials;Commercialization;Cryopreservation;Culicidae;Cyclic Gmp;Data;Development;Disease;Disease Transmission;Disorder Control;Dose;Egg;Elements;Embryo;Ensure;Equipment Failure;Europe;Feeding;Female;Food;Funding;Genetic;Genetic Drift;Genotype;Goals;Government;Growth;Hatching;Hepatocyte;Improved;Incubated;Individual;Infection;Institution;International;Intramuscular;Invaded;Larva;Licensure;Life;Liver;Malaria;Malaria Vaccines;Manufacturing Process;Methodology;Methods;Natural Regeneration;Needles;Netherlands;Nih Vaccine Research Center;Nonhuman Primate;Novel;Offspring;Partner In Relationship;Pathogen;Pharmaceutical Preparations;Phase;Phenotype;Plasmodium Falciparum;Prevent;Product Development;Production;Programs;Prophylactic Treatment;Public Health Relevance;Radiation;Regimen;Reliance;Repository;Research;Risk;Route;Salivary Glands;Scale Up;Security;Small Business Innovation Research Grant;Source;Sporozoite Vaccine;Sporozoites;Staging;Success;Syringes;Technology;Testing;Time;Timeline;Tool;United States National Institutes Of Health;Vaccination;Vaccine Evaluation;Vaccine Research;Vaccines;Volunteer;Water Quality;Whole Organism;Work;