Oncodetect, Inc. 

In quest for superior diagnosis and treatment


Innovative diagnosis and therapeutic strategies

HomeDevelopment of recombinant p21 protein as a viable therapy for AIDSINVESTORS NEEDEDSCIENCE CONSULTANTNEWSExecutive SummaryBreast Cancer testp21 drug development

Company Overview

The company will utilize proprietary and patent-pending technology in the field of breast cancer detection, monitoring transplant rejection and the development of drugs to treat organ transplant recipients, cancer and heart patients.

 OncoDetect Inc. is a biopharmaceutical company focused to develop molecular diagnostic assays for the pro-diagnosis, diagnosis, and management of breast cancer and organ transplantation. The company is also dedicated to the discovery, development and commercialization of novel, small molecule drugs for the treatment of cancer, allograft rejection, and atherosclerosis.

The business will seek to commercialize:

    A blood-based diagnostic assay that indicates the presence or likelihood of breast cancer through measurement of molecular/protein markers that are uniquely expressed in malignant tumors.

    The therapeutic use of a p21-based protein to suppress malignant proliferation of cancer cells, inhibit the allo-immune activation that results in transplant rejection, and limit the progression of arteriosclerosis.

    A blood-based diagnostic assay to monitor the success (acute rejection, nephrotoxicity, or chronic rejection) of renal transplants.

    Breast Cancer Diagnostics:


     Current Diagnostic Techniques:

     A National Cancer Institute (NCI) report estimates that about 1 in 8 women in the United States (approximately 13.3 percent) will develop breast cancer during her lifetime. The American Cancer Society estimated that in 2003, 211,300 new cases of invasive breast cancer were diagnosed, along with an additional 55,700 cases of in-situ breast cancer. In 2004, 41,800 women were expected to have died from breast cancer (2nd only to lung cancer for cancerous deaths).

    X-ray mammography is by far the most prevalent diagnostic technique. Ultrasound and MRI are used when x-ray mammography is likely to be inconclusive because of highly dense breasts. Sensitivity of mammography is high, ~90%, but specificity is poor, ~40% (leading to biopsies in ~50% of overall cases that turn out to be benign).

     Digital mammography and computer-aided detection is gaining ground only slowly (the American Cancer Society lists 300 machines in the US) due to high cost and limited gains in effectiveness.

     Experimental noninvasive techniques include magnetic resonance elastography, PET scans, electrical impedance spectral imaging, microwave imaging spectroscopy, near infrared spectral imaging, and terahertz pulse imaging.

     Biopsy is required when a breast image reveals a suspicious lesion. Between 800,000 and 1.25 million biopsies are performed each year with a cost of $800 to $3,000, depending on the type of biopsy performed. (USFDA Center for Devices – Radiological Heath & Office of Women’s Health). According to the American Cancer Society, approximately 80% of breast biopsies are benign. Since biopsies are generally performed when a breast image reveals a suspicious lesion, sensitivity is not applicable, but specificity is biopsies is excellent (~100%?).

     Many molecular and genetic tests are under development, but only one test is widely accepted: if a woman carries a hereditary mutation in genes BRCA1 or BRCA2, she has a 36%-85% chance of developing cancer before age 70 (National Cancer Institute). However, this test is of limited value since only a small percentage of breast cancers are hereditary in nature (to be reviewed).

     Will need to assess for new multiplex PRC assay the specificity and sensitivity compared to conventional tests (mammography and biopsy) and evaluate as appropriate for different types of cancer (DCIS, fibrosis and benign). If a new test can show greater sensitivity than mammography, it could replace mammography. If sensitivity is not high, but specificity is good, it could be used in conjunction with mammography to eliminate unnecessary biopsies.

     Breast Cancer Markers:

    In breast cancer cells, some genes are overexpressed while others are under expressed; the associated mRNA can be measured using polymerase chain reaction (PCR) techniques and overexpressed proteins can be measured using ELISA and Western blot analysis techniques. OncoDetect Inc. will develop diagnostic assay kits to accurately diagnose the presence and/or likelihood of breast cancer by determining the unique pattern of expression using the following molecular/protein markers. These markers are particularly useful because they can be measured non-invasively, via blood lymphocytes rather than biopsy.

    Mammaglobin B (protein) – positively correlated ; Skp2 (mRNA) – positively correlated; PPAR-alpha (mRNA) – positively correlated; PPAR- gamma (mRNA) – negatively correlated; TIMP-2 (mRNA) – negatively correlated; TNF-alpha ( mRNA) – positively correlated); Cycline E (protein or mRNA?) – positively correlated; MMP-9 (mRNA) – positively correlated; MMP-2 (mRNA) – negatively correlated (Figure below)

     Research to Date:

    Research by Dr. Khanna has demonstrated the following results significant to the development of breast cancer diagnostic assays:

     Application of standard PCR techniques to malignant and non-malignant tissues from 50 human breast tissue pairs has confirmed the statistically significant difference in expression of molecular and protein markers between diseased and non-diseased states. (Results for Mammaglobin B, PPAR-alpha, PPAR-gamma, MMP-9, Skp2, TNF-alpha and TIMP-2.)

     Correlation of Mammaglobin B in breast tissues and lymphocytes of patients known to have malignant breast cancer (based on 25 tissue/lymphocyte pairs) indicates that mammaglobin B levels can be detected in blood samples, not only tissue samples requiring biopsy.

     Low concentrations of Mammaglobin B in the lymphocytes can be amplified using 10 to 40 PCR cycles, greatly lowering the detection threshold.

     Correlation of multiple molecular markers with in tissue and lymphocytes (done for a single patient, to be completed for remaining sample of 25 patients for which complete samples exist).

     Intellectual Property Development:

    Development of intellectual property for the breast cancer diagnostics will focus on: This is based on our PATENT APPLICATION "Patterns of gene expression and uses therefeof for the detection and diagnosis of breast cancer. US PROVISIONAL PATENT APPLICATION No. 60/642,481"

    Diagnostic kit using multiplex PCR assay(s) to diagnose breast cancer and assess genetic predisposition toward breast cancer.

     Multiplex PCR assays that includes a combination of genetic markers that have been statistically selected to maximize the selectivity and specificity of the test

     Diagnostic kit that includes a combination of PCR assays where one assay is comprised of genetic markers that increase with the likelihood and/or presence of breast cancer and another assay is comprised of genetic markers that decrease with the likelihood and/or presence of breast cancer

     Diagnostic kit using ELISA techniques to diagnose presence of breast cancer based on increased/decreased expression of proteins

     Diagnostic kit based on applying multiplex PCR to blood lymphocytes for the purpose of diagnosing breast cancer and/or assessing the genetic predisposition toward breast cancer

     Diagnostic kit based on applying multiples PCR to nipple discharge and/or ductile lavage

     Unique primers used in development of PCR assays for specific genetic markers

     Development Milestones:

    Select best target markers for multiplex assay (part of phase I feasibility testing)

    Submit SBIR Phase II grant(s) for prototyping of breast cancer diagnostic assay

    Develop real-time multiplex PCR assay using best candidate markers (part of Phase II prototyping)

    Phase III commercialization will include: development of diagnostic kit and quality system.

    Determine FDA regulatory strategy and submit appropriate documentation – 510k submission (market to labs as research product) or PMA (requiring additional testing allowing product to be marketed as diagnostic product)

      p21 as a potent immunosuppressive drug:

    Thes studies demonstrate that by virtue of its potent anti-proliferative effects, cyclin kinase inhibitor p21 can render host lymphocytes less responsive to allo- and mitogenic stimuli; induce tolerance and prevent alloimmune activation enabling a prolonged graft survival. Additionally, its known inhibitory effects on tumor cells, protective effects on renal function and abrogation of viral replication will allow p21 to be an attractive alternate strategy to achieve immunosuppression for organ transplant recipients without side effects associated with current immunosuppressive drugs.

    Recombinant p21 protein when injected to mice results in the inhibition of lymphocyte proliferation and markers of inflammation and does not cause any toxicity Western blot analysis has confirmed that p21 enters cell nucleus within 4 hours interact with transcription factors and inhibits gene and protein expression of proinflammatory cytokines.Recombinant p21 protein inhibits lymphocyte proliferation similar to the currently clinically used immunosuppressive agents; Cyclosporine, Tacrolimus and Sirolimus Overexpression of p21 in recipients of rat cardiac transplants prolongs graft survival without immunosuppressive agents by inhibiting intragraft expression of pro-inflammatory cytokines.

    Intellectual Property Development:

    Development of intellectual property for the p21 based therapies will focus on: Intellectual Property Development: Development of intellectual property for the P21 based therapies will focus on based on our Patent application" Cyclin inhibitor p21: potent inhibitor of cellular proliferation; potential therapeutic applications in Transplantation, Cancer and atherosclerosis. US PROVISIONAL PATENT APPLICATION No. 60/638,930"

    Use of p21 as a therapeutic to address abnormal cell proliferation

    Use of p21 as an anti-proliferation applied to cancer, transplant rejection and arteriosclerosis

    Use of the protein transducing domain properties of p21 that allow p21 to penetrate the cell cytoplasm and the cell nucleus without the cumbersome process of attaching the appropriate protein transducing domain that is required to utilize other cyclin kinase inhibitors such as p16, p23, p27, p53, p73

    Use of p21 with targeted mutations to selectively increase or decrease cell proliferation by creating proteins that can enter

    The prototyping phase of development will include statistical analysis to examine which combination of markers provides the best diagnostic results with respect to sensitivity and specificity

Components of blood based molecular diagnostic assay for breast cancer. Real Time PCR analysis indicate similar mRNA expression of mammaglobin B in breast tissue (T) and Lymphocytes; mRNA expression of Skp2 and MMP-9 only in malignant but not in normal tissue from the same breast. Significantly different expression in breast tissues from patients and controls is also shown.