SBIR-STTR Award

Chronic skin wounds associated with various diseases (e.g., diabetes) and aberrant healing from acute wounding (e.g., hypertrophic scarring) is a major health care burden, which need scientific discovery-based therapeutic interventions. Our previous studies show that Smad7, a TGFb signaling antagonist, accelerates skin wound healing. We have developed a Smad7 fusion protein that contains the human Smad7 fused to the HIV-1 Tat protein transduction domain (PTD). The Tat-Smad7 protein can rapidly penetrate cells upon contact. Our preliminary data revealed that topical Tat-Smad7 application to skin wounds accelerated healing in wildtype and diabetes (db/db) mice. Our preliminary data also suggest that the Tat fusion protein containing either the N-terminal 258aa of Smad7 (Tat-N-Smad7) or the C-terminal 259-426aa of Smad7 (Tat-C-Smad7) could have full or partial effects of Tat-Smad7. This Phase I STTR application proposes to compare fusion protein efficacies of full length Smad7 (Tat-Smad7), N-terminal Smad7 (Tat-N-Smad7) and C-terminal Smad7 (Tat-C-Smad7) on wound closure in vitro and in vivo, and establish quantification methods for quality control of their bioactivities for future commercial use. Aim 1 will produce full length Tat-Smad7, Tat-N-Smad7 and Tat-C-Smad7, and determine if quantification of keratinocyte migration and proliferation can be used for quality control among different batches of Tat-Smad7 and its truncated derivatives. Cultured human keratinocytes will be treated with these proteins and quantify their effects on keratinocytes proliferation and migration via live-cell imaging. We will also stain nuclear pSmad2 and NFkB p50 to determine if their biological effects are associated with blocking TGFb and NFkB signaling as seen in full length Tat-Smad7. Aim 2 will compare the efficacies of Tat-Smad7 and its truncated derivatives on wound healing in vivo. We will test the 3 Tat-Smad7 variants on excisional wounds in normal and db/db mice to compare the rates of wound closure and re-epithelialization during healing and fibrotic response during wound remodeling. Tur proposed studies will narrow the lead Smad7-based Tat fusion protein(s) to further develop into topically applied therapeutic biologics to treat skin wounds. Completing this application will prepare us to perform IND-enabling studies via a Phase II application related to formulation and long-term toxicities of these biologics in vivo, as well as pig wound studies using GMP grade recombinant proteins.

Public Health Relevance Statement:


Public Health Relevance:
Chronic skin wounds affect 6.5 million patients in the United States. This application test novel biologics that can treat chronic or scarring skin wounds by topical applications.

NIH Spending Category:
Diabetes

Project Terms:
Acute; Affect; base; Biological; C-terminal; care burden; cell motility; Cells; cellular imaging; Chimeric Proteins; Chronic; Cicatrix; commercialization; comparative efficacy; cost effective; Data; db/db mouse; Diabetes Mellitus; Disease; DNA Binding; Drug Formulations; Escherichia coli; Exhibits; Family suidae; Fibrosis; Future; Goals; Healed; healing; Healthcare; HIV-1; Human; Hypertrophic Cicatrix; In Vitro; in vivo; Infection; Inflammation; keratinocyte; Lead; Length; Life; Link; MAP Kinase Gene; Mediating; Methods; migration; Molecular; Mus; N-terminal; NF-kappa B; Non-Insulin-Dependent Diabetes Mellitus; novel; Nuclear; oral mucositis; Oral mucous membrane structure; Patients; Peptides; Phase; Physiological; prevent; Production; prophylactic; Proteins; public health relevance; Quality Control; Radiation; Recombinant Proteins; research study; response; Signal Transduction; Skin; Smad7 protein; Small Business Technology Transfer Research; Splint Device; Staining method; Stains; Surgical wound; tat Protein; Testing; Therapeutic; Therapeutic Effect; Therapeutic Intervention; TNFRSF5 gene; Topical application; Toxic effect; Transactivation; United States; Variant; wound; Wound Healing

Chronic skin wounds associated with various diseases (e.g., diabetes) and aberrant healing from acute wounding (e.g., hypertrophic scarring) are a major health care burden. This health burden increases in aging populations. The cost to treat these conditions exceeds $25 billion annually. Currently, REGRANEX®, a cream containing a biologic based on human PDGF (Platelet-Derived Growth Factor), is the only FDA (Food and Drug Administration) approved biologic to treat diabetic wounds. However, the combination of its modest effect on healing and a black box warning added by the FDA for its cancer risk have not translated its drug efficacy to clinical benefit. These facts highlight the sense of urgency for scientific discovery-based therapeutic interventions. Allander Biotechnologies is developing proprietary topically applied biologics to promote wound healing. During the Phase I funding period of this grant, we have tested several biologics and identified our lead biologic to be further developed in this Phase II application. Our Phase I studies revealed that our biologic has multiple functions needed for wound healing. Our Phase II studies will focus on the process of formulation development for our biologic to be used as a topical drug, and data production that will lead us to a successful path for filing an Investigational New Drug (IND) application to the FDA. We will perform pre-formulation studies for solubility as well as stability of physicochemical characterizations and bioactivities of our drug substance. We will use data generated from pre-formulation studies to guide our formulation prototype development. We will produce our biologic at pharmaceutical grade purity and use it as well as the lead formulation(s) to treat diabetic mouse wounds and pig excisional wounds for efficacy studies to define pharmacodynamics (PD) biomarkers and identify potential acute cutaneous/systemic toxicities. By the end of this Phase II funding, we will have stability and physicochemical profiles of our biologic, the lead formulation, PD biomarkers, and protocols (analytical, bioassay, and toxicology) standardized for generating additional IND data under Good Laboratory Practice conditions.

Public Health Relevance Statement:
NARRATIVE Chronic skin wounds in diabetic patients and the elderly, affect 6.5 million people in the United States. This application aims to develop a novel biologic that can treat chronic and acute skin wounds by topical application to alleviate immediate suffering caused by skin wounds and prevent long-term wound related sequelae, such as permanent tissue/organ loss or scarring.

NIH Spending Category:
Dental/Oral and Craniofacial Disease; Diabetes

Project Terms:
Acute; Affect; Aging; Animals; Area; Autopsy; base; Biological Assay; Biotechnology; Blood Circulation; Buffers; cancer risk; care burden; Cells; Chronic; chronic wound; Cicatrix; Clinical; Collaborations; cost; Cream; Cutaneous; Data; design; Development; Diabetes Mellitus; Diabetic mouse; diabetic patient; Diabetic wound; Disease; Dose; drug development; drug efficacy; Drug Kinetics; efficacy study; Elderly; Epidermal Growth Factor; Excipients; Excision; Family suidae; FGF2 gene; Formulation; Funding; good laboratory practice; Grant; Gray unit of radiation dose; Growth Factor; Guidelines; healing; Health; Healthcare; HIV-1; Human; Hypertrophic Cicatrix; Inflammation; Investigational Drugs; Investigational New Drug Application; Ionic Strengths; keratinocyte growth factor; Lead; Length; Maximum Tolerated Dose; Measurement; Modeling; Mus; novel; Organ; Patients; Penetration; Peptides; Pharmaceutical Preparations; pharmacodynamic biomarker; Pharmacodynamics; Pharmacologic Substance; Phase; phase 1 study; phase 2 study; Phase I Clinical Trials; Platelet-Derived Growth Factor; Population; Preclinical Testing; prevent; Process; Production; Property; Proteins; Protocols documentation; prototype; Research Contracts; response; Rodent; safety study; Sampling; scale up; Serologic tests; Serum; Signal Transduction; Skin; Skin wound; Solubility; Standardization; Study models; systemic toxicity; tat Protein; Testing; Texture; Therapeutic Intervention; Tissue Sample; Tissues; Topical application; Toxic effect; Toxicology; Transforming Growth Factor alpha; United States; United States Food and Drug Administration; Variant; wound; wound closure; Wound Healing