Comosain Alternative Treatment
Three Phases of Experiments
Phase I: Tumor cell lines experiment
6 cancer cell lines were used for this research. Cancer cell lines from breast, lung, colon, ovarian, cervical and uterine origins were cultured and Comosain in an amount of 0.2, 0.4, 0.6, 0.8, 1.0mg/ml concentration were administered in cell cultural solutions. The result are outstanding: 50% of all cancer cell lines were eliminated in the concentration of Comosain in amount of 0.4 to 0.6 mg/ml, and more than 90% of cancer cell lines were eliminated at the concentration of 1mg/ml.
Phase II: Animal Experiment(Example 3)
Comosain inhibits cancer cell growth in vivo on experiemntal animal.
Comosain intraperitoneal administration to experimental animal.
14 experimental animals of 4 to 6 week old white rabbits each weighting 1 to 1½ pounds were fed under a condition of 23+_3 degree C, relative humidity of 45+_5 % and photoperiod of 12 light/12 dark. The rabbits were divided into seven groups of 2 heads each and were fed with Harlan-Taklad rabbit diet TD-1376 containing moisture 12%, crude protein 16%, crude fat 2%, crude fiber 15% , ash 8%, and 47% of nitrogen free substances.
The rabbits were fed for 3 weeks with free access to the diet and water. Body weight was recorded every 7 days, and records were analyzed. All rabbits showed a normal growth rate with no significant differences among the 7 groups in regard to the diet ingestion amount or the body weight gain. The cancer cell lines were injected into six groups of the rabbits, that is 2 heads each group with 2 heads serving as controls (without tumor cell injection). The cancer cell lines were developed from Example-2. Each head was injected 0.5ml of different cell line fluid intraperitoneally, prefer in the peritoneum layer, then the rabbits were fed the same diet for 3-4 weeks until a tumor grew in the peritoneum. The size and location of the tumors were recorded. When the tumors reached 3-5mm diameter in size, Comosain in the amount of 25.0mg/ml in normal saline with 100mg of vitamin C (to keep the solution acidified), one ml of Comosain was injected into the six different group of rabbits, the Comosain were given twice a week for 8 weeks.
After 8 weeks of treatment, the rabbits were anesthetized with injections of ketamine 75mg/kg in the femoral muscle and sacrificed. Blood samples were collected from the heart of each rabbit to determine the blood analysis which consisted of: complete blood count (CBC), Chemistry-7 and 24, (including liver and renal function tests), lipid profiles (including total cholesterol, HDL, LDL, VLDL, and triglycerides), coagulation factors consisting of; prothrombintime (PT), partial thromboplastin time (PPT), and immune-globulin –E. All the laboratory tests were analyzed, and indicated no differences among or within each groups. All laboratory blood analyses were performed on rabbits of all 7 groups. The results were tested using student t-test and Microsoft Excel-7 programs. The results are depicted in Table-II.
Table–II, The table presents blood analysis of the rabbits of 6 different groups which were treated with Comosain after inoculation of cancer cell lines. Control group; TC (183.3+_50.2mg/dl ), TRG (110+_40.6mg/dl), HDL (45.6+_20.4mg/dl), SGOT (38.6+_6.2u/l), SGPT (62.5 +_6.5u/l), GGTP (8+_ 2.4u/l ), WBC (6.8 +_ 2.0k/ul), Hb; (12.3+_2.2gm/dl). Bromelainase treated groups; TC (175.6+_ 36.8mg/dl), TRG (92.6 +_ 38.8mg/dl), HDL (43.6 +_16.5mg/dl), SGOT (110.8 +_30.7u/l ), SGPT (71.2 +_3.8u/l ), GGTP (7+_1), WBC (7.3+_ 2.2k/ul), Hb (11.9+_1.9gm/dl). TC; Total Cholesterol, TRG: Triglycerides, WBC: White Blood Cell, HDL; High Density Lipoprotein, SGOT; Serum Glutamo-Oxalic Transferase, SGPT; Serum Glutamo-PyruvicTransferase, Hb; Hemoglobin.
The internal organs from the rabbits sacrified in this sample including lung, heart, liver, kidney, muscle, omentum, intestine, stomach bladder and pancreas were visual examined and showed no abnormalities. One half of each organ was frozen, and the other half was fixed with 10% neutral buffered formalin for 24 hours. Then the fixed organs were washed with tap waterand stepwise dehydrated with 70%, 80%, 90% and 100% ethanol and then embedded in paraffin using Shandon-Histocentre-2. The embedded organ blocks were sectioned in 4mm thickness with a microtome (McBain, M 820, American Optical Co. USA) and stained with hematoxylin and eosin stain (H.E. stain). The stained specimens were made transparent with xylene, and mounted with permount on microslides. There were no pathological abnormalities or lesions under microscopic examination. All the specimens collected from six group of rabbits showed no evidence of persistent disease or cancer cells.
Therefore, we concluded that Comosain can served as a potent chemotherapeutic agent in various types of cancer treatment in experimental animal without side effects.
Phase III: Human Volunteer Experiment (Example 4)
Comosain oral administration to inhibit tumor growth in humans.
Comosain oral administration to late stage cancer patients.
Twenty-four volunteers were divided into six groups (four in each group) and 6 persons served as a control group (no Bromelain treatment). All were in their 4th and 6th decades with various types of cancers including breast, lung, colon, ovarian, cervical and uterine origins. All were in either Stage III or Stage IV. (The cancers had metastasized widely either to lung, liver, bladder or rectum). All had been treated with either radiation or chemotherapy after surgery but experienced no positive results. The Bromelain was administered in doses of 50-60 mg/kg based on 50-60kg of body weight. That is 2,000 to 3,000 mg/day divided into two doses.
Patients were monitored with bi-weekly blood tests; consisting complete blood counts, chemistry-7, chemistry-24, kidney and liver function tests, tumor markers, coagulation factors, and X-ray or CT scan in appropriate areas to determine the size of the tumors. There were no abnormalities in all the blood tests, no anemia, no leucopenia, no thromcytopenia, no abnormal kidney nor liver function tests. Tumor markers decreased; and tumors shrunk in size on the X-ray or CT scan measurement. Patients’ lifestyles become manageable and improved considerably. The treatment periods varied from 6 to 9 months. At this report no patients in the treatment group have expired. However, all patients in control group, who had not wished to be treated, succumbed to their related cancers in 6-12 months.
(Step-A) After blood samples were collected and allowed to stand for 2 hours, they were centrifuged at 4000rpm for 10 minutes. The superantants were separated and stored in a deep freeze before analysis. The chemistry analysis was carried out by blood chemistry analyzer to determine the changes in total cholesterol, HDL, LDL, triglycerides, liver function tests (such as SGOT, SGPT, G-GPT), renal function tests, and coagulation factors (PT, PTT). All results were tested with student t-test and Microsoft Excel-7.0 program. The results are depicted in Table-III, which depicts the blood analysis of 24 volunteers suffering from various types of cancer with Comosain oral therapy.
TC; (210.3+_30.2mg/dl ), TRG; (165.5 +_ 28.3mg/dl), HDL; (43.3 +_ 22.2mg/dl), SGOT ; (34.7+_6.2 u/l), SGPT; (63.3+_5.6 u/l), GGTP; (7.2+_ 2.1 u/l) WBC; (6.7 +_ 2.8 k/ul), Hb; (12.3+_ 2.1gm/dl)
Therefore, we concluded that the treatment of these various represented types of cancers for prolonged periods with BROMELAIN (COMOSAIN ) are effective and without side effects.