BIRM Estudios

CIEN T Í FICA MEN T E CO M P RO B A D O

Estudio de la Universidad de MIAMI, 2016

BIRM provoca apoptosis (muerte programada) de células cancerígenas.

ABSTRACT

BIRM is an anticancer herbal formulation from Ecuador.

Previous study established its antitumor and antimetastatic activity against prostate cancer models. The activity of BIRM against human prostate cancer (PCa) cells was investigated to uncover its mechanism of antitumor activity. In androgen receptor (AR)-expressing PCa cells BIRM was 2.5-fold (250%) more cytotoxic in presence of androgen (DHT) compared to cells grown in the absence of DHT. In AR-positive cells (LAPC-4 and LNCaP) BIRM caused a dose and time-dependent down-regulation of AR and increased apoptosis.

Exposing cells to BIRM did not affect the synthesis of AR and AR promoter activity but increased degradation of AR via proteasome-pathway. BIRM caused destabilization of HSP90-AR association in LAPC-4 cells. It induced apoptosis in PCa cells by activation of caspase-8 via death receptor and FADD-mediated pathways. A synthetic inhibitor of Caspase-8 cleavage (IETD-CHO) aborted BIRM–induced apoptosis.

The effect of BIRM on AKT-mediated survival pathway in both AR+ and AR- negative (PC-3 and DU145) showed decreased levels of p-AKTser 473 in all PCa cell lines. BIRM dosed by oral gavage in mice bearing PC-3ML tumors showed selective ef cacy on tumor growth; before tumors are established but limited ef cacy when treated on existing tumors.

Moreover, BIRM inhibited the LNCaP tumor generated by orthotropic implantation into dorsal prostate of nude mice. Partial puri cation of BIRM by liquid-liquid extraction and further fractionation by HPLC showed 4-fold increased speci c activity on PCa cells. These results demonstrate a mechanistic basis of anti-tumor activity of the herbal extract BIRM.

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Estudio de la Universidad

Maharaja Sayajirao

de India, 2016

BIRM como analgésico y reductor del dolor neuropático

ABSTRACT

Neuropathic pain condition remains poorly managed by currently available therapeutics.

There is therefore a dire need for development of efficacious therapeutics with minimal side effects. BIRM , an extract of Amazonian plan solanum dulcamara, consumed as a dietary supplement by natives in Ecuador, is considered as a natural remedy for a number of ailments (AIDS and Cancer, among others). The aim of the current study was to test the efficacy of BIRM in in vivo neuropathic pain model to elucidate its anti-neuroinflammatory potential. Rats subjected to chronic constriction injury (CCI) were divided into CC-control, CCI-Gabapentin and CC-BIRM group along with a normal control group.

BIRM was administered orally (4 ml/kg, daily) to animals group from day 14 post surgery till day 28. Repeated oral administration of BIRM inhibited CCI-induced mechanical allodynia and thermalhyperalgesia. It also inhibited CCI-induced activation of microglial cells and upregulation of COX-2 and TFN-α in the dorsal horn of the lumbar spinal cord. These data indicate that the marketed formulation BIRM, has anti-neuroinflammatory and anti-nociceptive properties in neuropathic rats and can serve as an adjuvant to standard therapy or as a stand-alone therapeutic agent for the treatment of neuropathic pain disorders.

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CANCER

PREVENTION & TREATMENT Miami 2014 Dr. Cevallos Speech:

Good Moming! It is indeed a pleasure to Frescya and l to be present at the Inaugural University of Miami Cancer Symposium. I was very pleased to meet with the distinguished speakers and i look forward to listening to their talks. I am also looking forward to student presentations, because Frescya and I aIways believe that we must motivate the younger generation to choose the path of translational research, especially in areas of cancer chemoprevention and treatment.

In my clinical practice and academic pursuits l have always been interested in cancer chemoprevention and treatment of late stage cancers. These efforts became a reality when l formulated BIRM, a herbal supplement and founded Ecua BIRM Co. Although for many years in my clinical practice, l have successfully treated early and late stage cancer patients, and BIRM is consumed in more than 28 countries all around the world, I have always been interested in understanding the molecular mechanisms of why it works. About ten years ago, I got that opportunity. An elderly patient of Dr. Mark Soloway's who had developed bone metastatic prostate cancer began showing improvement in symptoms and a decrease in serum PSA levels.

S0 Dr. Soloway asked him whether he was drinking any magic potion. Sure enough, a week later, the patient brought him a bottle of BIRM. And as the story goes, one late at night, Dr. Soloway called Dr. Bal Lokeshwar and asked him whether he could cure prostate cancer in his rats by feeding them BIRM. This was the beginning of a long collaboration. Drs. Bal and Vinata’s results showed that indeed BIRM had potent cancer chemopreventive and treatment efficacy in several different models of prostate cancer and it could inhibit metastasis promoting enzymes. Through long and hard fought efforts, Bal was able to obtain a 5-year NIH grant to identify the molecular mechanisms of the anti-tumor and anti-metastatic properties in BIRM. For this grant, working in collaboration with my daughter Ms. Bonny Cevallos, Ecua BIRM Co, fully and confidentially disclosed to the NIH the scientific method of preparing BlRM. So what happened to the patient, Mr. Degetau peacefully passed away a few years ago due to cardiovascular disease and not prostate cancer.

Dr. Lokeshwar wants me to tell you that neither Bal, Vinata nor Dr. Soloway have received any financial benefits personally or professionally from us or from Ecua BIRM. But this story I hope shows you how a good collaboration works. To honor this collaboration we decided to support the University of Miami Cancer Symposium. Since it all began with Dr. Soloway, who is passionate about research, we wanted to honor him. Therefore, at this time, I would like to invite honorable Dean Dr. Goldschmidt to join me in presenting this award to Dr. Mark Soloway, for his distinguished contribution to Urologic oncology research and patient care.

UNIVERSITY OF MIAMI CANCER SYMPOSIUM Cancer Prevention & Treatment PROGRAM 10.00 am - 1.00 pm Venue: 10.00 - 10.15 am

10.20 - 11.00 am

Title 11.00 - 11.40 am Title 11.40 - 12.00 pm 12.00 - 12.15 pm 12.15 - 12.55 pm Title 12.55 -1.10 pm 1.10 - 1.15 pm 1.15 - 2.45 pm Venue 2:45 pm

Session I: Cancer Chemoprevention & Treatment 7th Floor Lois Pope Auditorium Welcome & Introductory Remarks Pascal Goidschmidt, M.D. Senior Vice President for Medical Affairs and Dean Stephen D. Nimer, M.D. Director, Sytvester Comprehensive Cancer Center Edwin Cevallos, M.D. Founder & CEO Ecua BIRM Inc. William G. Nelson, M.D. Ph.D Marion I. Knott Professor and Director Sidney Kimmel Comprehensive Cancer Center Johns Hopkins University, School of Medicine Can Prostate Cancer Be Prevented?

Dr. Shrikant Anant, Ph.D Tom and Teresa Walsh Professor of Cancer & Associate Dean for Research University of Kansas Medical Center Cancer Stemness: Quiet Cells Need Knocking Down a Notch Coffee Break & Mixer with speakers Poster Award Winner Presenter 1 Victoria L. Seewaldt, M.D. Professor of Medicine Duke University In Praise Of Small Science: A Duke-Durham Partnership to Investigate the Biology of Interval Cancers Poster Award Winner Presenter 2 Concluding remarks Session II: Bag lunch and Poster Viewing Session Schoninger Quadrangle Announcement of all Poster Award Winners 4

BIRM

aprobado y

reconocido

internacionalmente La oficina de Patentes y Marcas de Estados Unidos luego de rigurosos anรกlisis y comprobaciones, otorgรณ a BIRM la patente Nยบ: US 7,250.180 B2 en la cual se reconoce la originalidad del producto autorizando su uso medicinal en la prevenciรณn y cura de varias enfermedades. Este certificado garantiza la calidad de BIRM y lo protege de cualquier intento de falsificaciรณn.

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UNITED STATES PATENT 2007

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Investigación

Induction of caspase-mediated apoptosis and Inhibition of Prostate Tumor Growth and Metastasis by a Plant extract BIRM.

del BIRM

Deven S. Dandekar1, Vinata B. Lokeshwar1, Edwin Cevallos.2, and Bal L. Lokeshwar1. ddandekar@miami.edu 1Dept. of Urology and Sylvester Cancer Center, Univ. of Miami.

Miami, FL. 2Tumor Institute, Quito, Ecuador.

en EEUU, 2003

Introduction: Most cancer drugs are derived from plants with many more to be discovered (1). An orally active, aqueous plant extract (Dulcamara sp) was tested for its anti-tumor properties on prostate cancer cells in vitro and a metastatic model in vivo.

Methods: Cytotoxicity of BIRM on three human and one rat-prostate tumor cell unes was evaluated by [3H]-Thymidine incorporation-inhibition and clonogenic survival assays. Apoptotic activity and caspase involvement was assayed by nucleosome release and fluorogenic caspase assays, respectively. Cell cycle-phase fractionation was determined by flow cytometry, Anti-hyaluronidase activity was assayed by an ELISA-Iike assay (2). Anti-tumor activity of BIRM was tested on the Dunning EGFP-MATLyLu rat tumor, a metastatic CaP model.

Results: BIRM inhibited cell proliferation (fig1) and clonogenic growth of CaP cell lines (IC 50: 0.8% ml/ml).

Cells treated with BIRM showed increased accumulation in G0/G1 phase (33.8%) and decrease

(54.6%) in S-phase. Cells incubated with BIRM underwent apoptosis via activation of caspase mediated cell death (Fig.2&3), BIRM inhibited activity of a tumor-derived hyaluronidase (IC50 -1 ml/ml). Oral gavaging with BIRM in MAT LyLu tumor-bearing rats caused a significant decrease in tumor incidence (50%), growth rate (1 cc tumor growth: 18.6 ± 1.3 days in control vs. 25.7 ± 2.6 days (38% growth delay) and a 63% reduction in lung metastasis (Fig.4&5).

Conclusion: Our results demonstrate that BIRM is a potent orally active anti-neoplastic agent, which primarily affeets caspase-mediated apoptosis and inhibits metastasis by inhibition of hyaluronidase, an ECM-degrading enzyme implicated in metastasis.

REFERENCES: 1. 2.Lokeshwar VB et al.

Cancer Res. 56:651-657, 1996.

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BIRM demuestra

cero toxicidad en los estudios

Birminghan, 1996

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BIRM en el Congreso

Mundial de SIDA Vancouver, 1996

BIRM: La estrategia terapéutica En base a los estudios presentados en la conferencia de SIDA en Japón en 1994, se presenta en el congreso mundial de SIDA en Vancouver, la actualización de resultados médicos: Los fenómenos inmunológicos y la apoptosis son controlados gracias al uso de oligomonosacáridos, nucleótidos y minerales obtenidos de la naturaleza. Experiencia multicéntrica de seis años 1989-1996. Cevallos Arellano Edwin, Mead of the Oncology Clinic of Metropolitan Hospital, Ardilla Ardilla Henry. Director of Liga Colombiana de Lucha contra el SIDA, Santa Fé de Bogotá., D.C. Colombia, Dr. Ignacio Mirazo Milillo. Director del Servicio de Enfermedades Infecto-contagiosas del Hospital Dr. José Scoseria. Montevideo Uruguay, Dr. Antonio Plaza. Director del Centro de Terapias Integrales, C.T.I., Madrid España., Dr. Juan Manuel Núñez Núñez. Director Médico del Centro de Medicina Natural, Tijuana México.

Objetivos: Ratificar la bondad del BIRM como estimulador y modulador del Sistema Inmunológico en pacientes con SIDA. Ratificar la ausencia de toxicidad del BIRM aún en altas concentraciones.

Ratificar la imposibilidad del virus para presentar resistencia al BIRM. Evaluar las propiedades estimulantes del Sistema Inmunológico, así como del efecto sobre la apoptosis, utilizando oligomonosacáridos y minerales de origen natural en 160 pacientes con SIDA. Métodos: 160 pacientes con diagnóstico confirmado de SIDA, son seleccionados de un total de 1000 pacientes, 40 colombianos, 40 ecuatorianos, 60 españoles, 10 uruguayos, 10 mexicanos para tratamiento oral con BIRM en períodos de 22 meses a 6 años.

En la evaluación fueron considerados síntomas clínicos y signos, porcentaje total de linfocitos CD4 y CD8 e índice. Los estudios in-Vitro han demostrado que BIRM controla al virus en un 88% y además impide la formación de syncytios en un 59%.

Resultados: Los resultados obtenidos en 22 meses y 6 años demostraron una mejoría clínica así como signos y síntomas en un 91.4% de pacientes, recuperación inmunológica en 94.3% y mejoría clínica e inmunológica en un 91%; 8 pacientes que representan el 5.7% murieron debido a su estado avanzado de enfermedad. Recibieron tratamiento durante un mes y los niveles de CD4 bajo 100.

Discusión y Conclusiones: El BIRM (Modulador Biológico de la Respuesta Inmune) ha demostrado ser el más potente estimulador y modulador del sistema inmunológico que se conoce. Compitiendo con los receptores CD4 y además evitaría el secuestro viral en las células dendríticas del ganglio linfático.

El BIRM es un viricida e impide la formación de masas sincitiales en un 59%.

Comparativamente con los nucleótidos conocidos reporta un 88% de control viral.

El BIRM por sus propiedades es necesariamente el tratamiento de elección en los pacientes con SIDA por carecer de toxicidad, por su fácil administración, por la imposibilidad del virus de desarrollar resistencia y por haber demostrado durante 6 años mejorar los niveles de CD4 en forma indefinida. Finalmente, el BIRM ha demostrado transformar una enfermedad necesariamente mortal en crónica.

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BIRM en la Décima

Conferencia Internacional

sobre SIDA Japón, 1994

PB0294 BINATIONAL EXPERIENCE IN THE TREATMENT OF AIDS WITH A LOW MOLECULAR WEIGHT NATURAL CARBOHIDRATE (ECA-10-142), AS STIMULATE OF THE INMUNOLOGY SISTEM, Cevallos Arellano Edwin, Head of the Oncology Clinic at the Metropolitan Hospital, Quito, Ecuador; Ardila Ardila, Henry, Director of the Liga Colombiana de Lucha Contra el SIDA, Santafé de Bogotá D.C., Colombia.

Objectives: To evaluate the stimulant properties of the inmnunological system of the (ECA-10-142) a low weight natural origen carbohidrate, in 70 IV CDC stage AIDS patients.

Methods: 70 patients with AIDS confirmed diagnosis were selected, 30 colombians and 40 ecuatorian, for oral treatment with (ECA-10-142). In a period of 9 months.

In the evaluations were considered clinical symptoms and signs, total percentage of linfocites, CD4 and CD8 cells percentage and its relationships.

In In vitro studies was demonstrated also that this carbohidrates stimulate the inmunological system in the AIDS pattients.

Results: The obtain results in the 9 months demonstrated a clinical improvement as to signs and symptoms in 64 (91.4%) patients, inmunological recuperations of the CD4 levels in 66 (94.3%) patients, clinical and inmunological improvement were presented in 64 (91.4%) patients, 4 (5.7%) patients died due to their advanced state of the sickness. They only received treatment during one month and had their CD4 levels under 100.

Discusion and Conclusion: The (ECA 10-142) has demonstrated to be an useful vitro and invivo stimulated of the inmunological system as inhibits the formation of sincitiales masses in a 59% and is efective as the clinical and inmunological improvernent of AIDS patients.

Kinetics studies are required and observations in a longer time in order to conclude it's inmunostimulating activity in AIDS patients.

PB0294 BIRM CARBOHIDRATE OF LOW MOLECULAR WEIGHT ECA 10-142 CONTROLS AIDS Cevallos A. Edwin Dr., Chief of Oncology and Radiotherapy Services of Metropolitan Hospital, Quito, Ecuador.

Objective: The BIRM ECA 10-142 in vitro, reports non-cytotoxic even at the hightest concentration tested, and moderated andti HIV activity with % reduction in syncitia-formation of 59%; (Southern Research Institute, Birmingham Alabama, U.S.A.; July 1991) In vivo double-blind reported an elevation in the number of CD4 cells.

The Chemical analysis report in Nuclear Magnetic Resonance stablished the empirical formula C3H502, or a multiple thereof.

Methods: Twenty patients in state IV of CDC, were chosen, and survived from january 1989 to april 1993.

In every patient the CD4 cells were elevated notoriously. For example: 149 CD4 cells to 480 CD4 cells, in 3 months of treatment with BIRM. Results: The results were evaluated based on the number of CD4, CD8 and quotient CD4/CD8; clinical evolution of the patient in his signs and symptoms. Patients with CD4 so low like 1%, survived for 15 months, and in the second group the patients are still alive after 22 months of treatment, Discussion and Conclussions: The BIRM ECA 10-142 has demonstrated been efective in all the states of HIV/AIDS, BIRM has been evaluated "in-vitro*"and "in-vivo"; and its composition has demonstrated to be an carbohydrated of low molecular weight, which permoms at the level of cellular adhesion and reduces in 59% the syncitia formation. The elevation in the number of CD4 cells indefinetly in all the patients treated, shows that we are in front of a modulater and stimulater substance of the Inmune System, a substance of enormous aplication in this kind of patients.

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Productos de la familia BIRM®

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