Myko San Research

Myko San Research

Myko San Research

Basic Research

Human Studies

Basic Research

Mushroom Extracts Against Fibrosarcoma, Squamous Cell Carcinoma and Melanoma

In 1999, Dr Myko San company initiated and in collaboration with the Ruđer Bošković Institute – Department for Molecular Medicine performed the first research of medicinal mushrooms in our country.

We have investigated in vitro antitumor effects of our single mushroom extracts from shiitake (Lentinus edodes), maitake (Grifola frondosa) and turkey tail mushroom (Trametes versicolor), and our blended mushroom extracts Lentifom and Lentram against very aggressive malignant mouse tumors fibrosarcoma, squamous cell carcinoma and melanoma. Effects were dose dependent; in optimal doses, single mushroom extracts and combined extracts showed very high tumor inhibition rates (up to 99.85%).

Importantly, the extracts (except T. versicolor) significantly stimulated the growth of healthy cells (fibroblasts).

Next, in preparation for the in vivo research on mice, we have performed the preliminary safety screening of single and combined mushroom extracts (lasting 42 days), which showed no toxic or any other harmful side effects in mice fed with the extracts.

In vivo research has shown that mice with advanced cancers (fibrosarcoma and squamous cell carcinoma) – treated with our mushroom extracts for 2 weeks – have greatly prolonged life span compared with control group. Adjusted for humans, this prolongation equals additional months and years of the human life.

Read more:

Ivanković S., Hiršl N., Jurin M. i Jakopović I.: The Influence of Medicinal Mushroom Preparations on Mouse Tumors. International Journal of Medicinal Mushrooms, Nr. 2/2004

Medicinal mushroom extracts inhibit squamous cell carcinoma and fibrosarcoma in culture (diagram)
The results of in vitro (on cell cultures of A) squamous cell carcinoma and B) fibrosarcoma) testing of Myko San extracts (CV = Coriolus versicolor (= Trametes versicolor), turkey tail mushroom; GF = Grifola frondosa, maitake; and LE = Lentinus edodes, shiitake). It is evident that each of these extracts strongly inhibit the proliferation of cancer cells, and the result is dose dependent – as dosage increases so does the inhibitory effect.
medicinal mushroom extracts improve survival of test animals (graph)
Laboratory mice were injected with squamous cell carcinoma on day 1 and their survival was tracked. At the conclusion of the study – day 34 – none of the mice in control group survived, while there was a 60% survival rate in the group treated with Grifola frondosa (maitake) extract by Myko San company.
Microscopy image of fibrosarcoma cells in culture left untreated vs. treated with medicinal mushroom extract by Myko San
Microscopy of fibrosarcoma cells growing in culture: on the left is the control (proliferating through the medium), on the right the same cells treated with 10% solution of Myko San’s mushroom extract Lentifom – only the remains of dead fibrosarcoma cells remain.

Anticancer Effects of 19 Single Mushroom Extracts and Combined Liquid Extract “Agarikon”

Following the excellent results of the first research, the Croatian Ministry of Education and Science sponsored our next research of antitumor effects of medicinal mushrooms, which was performed in cooperation with Bošković Institute – Department for Molecular Medicine. The project, lasting from 2006 to 2009, encompassed  in vitro tests of antitumor effects of 19 mushroom species (Cantharellus cibarius, Meripilus giganteus, Pleurotus ostreatus, Ganoderma lucidum, Fomes fomentarius, Hericium erinaceus, Grifola frondosa, Trametes versicolor, Agaricus blazei = brasiliensis = subrufescens, Calvatia gigantea, Poria cocos, Grifola umbellata, Lepista nuda, Piptoporus betulinus, Phellinus linteus, Stereum hirsutum, Tricholoma caligatum = matsutake, Schizophillum commune and Cortinarius violaceus) against extremely aggressive mouse fibrosarcoma and squamous cell carcinoma.

Nearly all single mushroom extracts elicited significant tumor growth inhibition; generally on both cancer cell lines and with larger concentrations resulting in stronger effect.

Additionally, we have tested the potential of our combined mushroom extract Agarikon on mouse breast cancer, colon cancer, squamous cell carcinoma and fibrosarcoma. Myko San’s Agarikon showed the strongest effects on all tested cancer cell lines. The effects were dose dependent – larger concentrations blocked cancer cell growth more effectively.

Agarikon.1 component strongly inhibits squamous cell carcinoma.
Effects of Agarikon.1 on squamous cell carcinoma in vitro.
Left is the control; on the right a 50% concentration of a single Agarikon.1 component demonstrating a direct cytotoxic effect (killing cancer cells).
Source: Rudjer Boskovic Institute
Graph showing that Agarikon strongly inhibits breast adenocarcinoma, bowel adenocarcinoma, squamous carcinoma and fibrosarcoma.
The effects of 25µL and 50µL of Agarikon medicinal mushroom extract on 4 tumor cell types (breast adenocarcinoma, bowel adenocarcinoma, squamous carcinoma and fibrosarcoma).

Anticancer Effects Mushroom Extracts: Blended vs. Single Species

(Tested on: human colon cancer, lung adenocarcinoma, small cell lung cancer, and brain astrocytoma)

In 2011, Myko San started, in cooperation with the Food and Biology Faculty (University of Zagreb), a new research project with the goal of solving a long-lasting dilemma regarding the size of antitumor effect in relation to the type of mushroom extract: single species extracts (or isolated compounds) and blended extracts.

The cytotoxic effects of Myko San’s blended mushroom extracts Lentifom, Super Polyporin,  Agarikon,  Agarikon Plus, Agarikon.1 and Mykoprotect.1 have been investigated on 4 human cancer cell lines (colon cancer, lung adenocarcinoma, small cell lung cancer and brain astrocytoma), and compared with popular simple products Beta Glucan and ImunoBran/MGN-3, as well as registered anticancer drug PSP.

All tested mushroom products showed cytotoxic effects on tested cancer types; they kill cancer cells by vitally damaging their cell membranes or mitochondria (the cell’s “power plant”, but also a major bio-signaling element that influences whether the cell will multiply or go to programmed cell death called apoptosis). The observed cytotoxic effects were dependent on cancer type and dosage, with better effects resulting from higher dosages.

Our blended mushroom extracts exhibited several times stronger antitumor effect on all tested human cancer types and in almost any applied concentration, compared with tested single mushroom extracts. In some tumor types, Myko San blended mushroom extracts (in the standard dosage) showed stronger antitumor effect than 100-fold dosages of single mushroom extracts.

Additionally, we have also measured the concentrations of total and soluble polysaccharides, and total polyphenols and total flavonoids. While mushroom polysaccharides (especially their beta glucans) are the main source of their strong antitumor and immunomodulatory properties, the activity of mushroom polyphenols and flavonoids – anti-oxidative, anti-mutagenic, anti-cancerogenic and anti-inflammatory – is also important. The interaction of polyphenols and flavonoids with high concentrations of polysaccharides especially improves their cytotoxic and immunomodulatory antitumor activity.

Finally, we have also tested the antitumor effects of 8 single mushroom extracts (Boletus regius, Cortinarius violaceus, Grifola frondosa – maitake, Meripilus giganteus, Craterellus cornucopioides, Pleurotus tuber-regium, Russula virescens, Scutiger pes-caprae) on 5 human cancer cell lines (colon cancer, lung adenocarcinoma, lung squamous cell carcinoma, small cell lung cancer and brain astrocytoma). Almost all of these extracts displayed promising antitumor effects.

Read more:

Durgo K., Končar M., Komes D., Belšćak-Cvitanović A., Franekić J., Jakopović I., Jakopović N., Jakopović B. Cytotoxicity of Blended Versus Single Medicinal Mushroom Extracts on Human Cancer Cell Lines: Contribution of Polyphenol and Polysaccharide Content. International Journal of Medicinal Mushrooms, Nr. 5/2013.

Agarikon.1 and Agarikon Plus Affect Cell Cycle and Induce Apoptosis in Human Colorectal and Non-Small Cell Lung Cancer

In 2013 we started, in cooperation with Biozyne (spin off company of Ruđer Bošković Institute), a research on mechanisms by which our blended medicinal mushroom extracts Agarikon.1 and Agarikon Plus influence cell cycle and induce apoptosis in human cancer cell lines (colon cancer and non-small cell lung cancer). Experimental methods included MTT proliferation assay, cell cycle analysis by flow cytometry, annexin V assay for detection of induced apoptosis and Western Blot analysis.

Both Myko San products exhibited significant antiproliferative (primarily cytostatic) action on both tested human cancers in concentrations of 1-10 mg/mL. They caused significant disturbance of cancer cell cycle in G1 and S phases, disturbing DNA replication in both cancer cell lines, which has been confirmed by increases in p53 and p21 protein expression.

Additionally, we found evidence of cancer cell apoptosis (programmed cell death) – both early and late – and necrosis in G1 phase of cell growth in both cancer types, as evidenced by Caspase-3 activation.

See the Scientific Presentation (7th International Mushroom conference, Beijing 2013)

Boris Jakopovich presentation
Boris Jakopovich presenting the findings of the study at the 7th International Medicinal Mushroom Conference in Beijing, 2013.

Human Studies

Human Study: Effects of Medicinal Mushroom Extracts in Patients with Bowel (Colorectal) and Breast Cancer

(presented at the 4th International Mushroom Conference, Ljubljana 2007)

Fifty-one patient with bowel (colorectal) cancer and 105 with breast cancer used intensive doses of Myko San products Lentifom, Super Polyporin and Agarikon (on average for 40-80 days), from the beginning of 2004 to mid-2007 at the same time or after their standard oncological therapies. Our analysis was based on the official medical records from hospitals where patients received the standard therapy and interviews with patients and/or their family members.

Bowel (colorectal) Cancer Patients (N=51)

At the beginning of mycotherapy (therapy using medicinal mushrooms), 60% of patients were in the most advanced stage 4, another 30% in stage 3.

At the end of intensive mycotherapy over 90% of patients had unchanged or improved status; 45% of total had improved status.

At the end of the research period (June 2007) 63% of patients were alive, with 80% of them experiencing improved or stable status (without disease progression).

Patients using our products noticed improved tolerance of chemotherapy and/or radiotherapy.

Data analysis revealed that intensive mycotherapy significantly improves the survival rate and health status, compared to standard therapy alone.

Breast Cancer Patients (N=105)

In the study 105 patients (103 women, and 2 men) with breast cancer (mostly ductal invasive carcinoma – 78 %) participated. 47% of patients had completely resected primary tumors, and 53% were in stage 4, the most advanced stage (metastases, recurrent cancer).

At the end of intensive mycotherapy 88% of patients had unchanged or improved status; 36% of the total had improved status.

At the end of the research period (June 2007) 61% of patients were alive. 69% of survivors became disease-free, 16% had improved or unchanged status, with disease progression in 15% of cases (recurrence rate: 3%).

Patients using our products noticed improved tolerance of chemotherapy and/or radiotherapy.

Data analysis revealed that intensive mycotherapy significantly improves the survival rate and health status, compared to standard therapy alone.

All of these results were much better than the results of standard therapy used at the time, as evidenced by official US cancer registers.

See the Presentation (given at the 4th International Mushroom conference, Ljubljana 2007)

Human Study: Effects of Medicinal Mushroom Extracts in Lung Cancer Patients

(presented at the 5th International Mushroom Conference, Nantong 2009)

Sixty-five patients with lung cancer (13 with small cell lung cancer and 52 with non-small cell lung cancer), had used intensive doses of Myko San products Lentifom, Super Polyporin and Agarikon (on average for 60-100 days), from the beginning of 2004 to mid-2007 concurrently or immediately following their standard oncological therapies. In June 2009, we collected the data from the official medical records and interviews with patients and/or their family members.

Patients with Small Cell Lung Cancer (N=13)

At the beginning of mycotherapy, nine patients were in limited stage and four in advanced stage.

At the end of intensive mycotherapy, three patients had complete regression, four had partial regression and one had progression (tumor size increase). The performance status remained unchanged in four and improved in three patients. Regarding chemotherapy, two patients experienced no side effects and five had alleviated side effects.

Since the cancer registers were not current in our country and the medical documentation was incomplete, we had to compare the median survival time of patients from first receiving our therapy (not from 1st diagnosis) with the US data (Skeel, Handbook of Cancer Chemotherapy, Lippincott, 2007) which measures survival time from the first diagnosis. Additionally, the survival rates at the time were higher in the US, than in Europe.

Even so, in limited SCLC, the US median survival time was just 14 months – compared to 37 months for patients treated with our products. For patients with advanced SCLC, the US median survival time was just 7-9 months – compared to 27 months for patients receiving intensive mycotherapy. We should remind again that the US figures count from the first diagnosis, while the mycotherapy treated group counts from the start of mycotherapy (approx. 6-12 months after the first diagnosis).

At the end of research (June 2009), four patients were alive with average survival time of 42 months. In May of 2015, two of them are alive and disease free (more than 9 years from the first diagnosis!).

Patients with Non-Small Cell Lung Cancer (N=52)

The sample with non-small cell lung cancer included 24 patients with adenocarcinoma, 3 large cell lung cancer, 13 squamous cell lung cancer and 12 non-specified cases. Seven patients had been operated and 45 had inoperable lung cancer (6 recurrent). Patients in the sample had very advanced lung cancer: 10 were in stage 3A, 20 in 3B and 20 in stage 4 (distant metastasis).

At the end of intensive mycotherapy, 15 patients had partial regression, 10 had no change and the disease progressed in three cases (measured in tumor size). The performance status remained unchanged in 12, improved in 10, and got worse in four cases. Two patients endured chemotherapy without side effects, nine with milder side effects and five had unchanged side effects.

At the end of the research period (June 2009), eight of patients were alive, and four of them were disease free.

Patients using our medicinal mushroom extracts lived longer and had better overall survival. As for small cell lung cancer sample, we have compared survival rates using US data (National Cancer Institute cancer statistics), measuring survival from the first diagnosis, with the medical data from our patients, measuring survival from the first administration of mycotherapy (usually 6-12 months after first diagnosis). Prolongation of life was evident for the whole sample of non-small cell lung cancer patients treated with medicinal mushroom extracts. In the most advanced stage 4 (metastatic NSCLC), for example, patients treated with intensive mycotherapy had 18% 4-year survival rate compared to just 2% for patients treated with standard oncological therapy alone.

Patients using our products also noticed better tolerance to chemotherapy and/or radiotherapy, significantly reducing their side effects.

Especially noteworthy was the clear dose-related impact of intensive mycotherapy on survival rate and improved health status for patients with NSCLC. For example, two years after the mycotherapy started, the group treated for 70-100 days had 40% survivors, while the group treated for longer (110+ days) had a 60% survival rate.

Patients who, following the initial intensive mycotherapeutic regimen, continued to use our medicinal mushroom extracts periodically enjoyed longer lives and increased survival in both small cell lung cancer and non-small cell lung cancer.

See the Presentation given at the 5th International Mushroom Conference, Nantong 2009

Human Study: Long Term Effects of Using Medicinal Mushroom Preparations in Human Colorectal and Breast Cancer

(presented at the 6th International Mushroom Conference, Zagreb 2011)

As a continuation of the study on effects of mycotherapy in patients with bowel and breast cancer, presented at the IMMC4 (Ljubljana 2007), this study included 52 patients with colorectal and 89 with breast cancer. They used our medicinal mushroom extracts Lentifom, Super Polyporin and Agarikon (for 40-270 days, 70 days on average), with periodical inclusion of Agarikon and Agarikon Plus following the initial intensive regimen. The patients started using intensive doses of our extracts during Jan/2005-Jan/2006 (either simultaneously or following their standard oncological therapies) and were followed until Dec/2010. We used official hospital records and interviews with patients and/or their family members.

Bowel (colorectal) Cancer Patients (N=52)

Our study focused mostly on advanced cases of colorectal cancer. At the beginning of mycotherapy 68% of patients were in stage 4 (the most advanced), and 26% in stage 3.

There were 27 patients alive with metastatic colorectal cancer at midpoint (August 2007), and 10 at the conclusion of the study (Dec/2010).

Metastatic colorectal cancer most often spreads to the liver and greatly decreases survival. Once metastases develop, chemotherapy can only reduce them in about 16% of cases, often with serious hepatotoxic side effects (like blue liver syndrome and steatohepatitis). In the subsample of bowel cancer patients with liver metastases, the intensive mycotherapy caused regression in 20% of cases, without any hepatotoxic side effects. (Just the opposite is true – available evidence suggests many medicinal mushrooms protect the liver.)

For colorectal cancer patients, US overall 5-year survival rate was 62% (National Cancer Institute and American Cancer Society data) – much better than 43% in Europe (European Journal of Cancer). Median survival for bowel cancer patients in USA was 29.2 months (from the first diagnosis), compared to 38 months in patients treated with our products (and measured from initial mycotherapy!).

Exceptional difference was found in patients with stage 4 bowel cancer (metastatic): 5-year survival in the USA using standard oncological therapy was just 5-8%, compared to astounding 26.5% survival rate in patients treated with Myko San medicinal mushroom extracts. (Again, the difference would have been even bigger if we had sufficient information to measure from the first diagnosis, and not from the start of our mycotherapeutic regimen!)

Breast Cancer Patients (N=89)

In our sample, 42% of patients had their primary tumor removed, while 56% were in stage 4 (most advanced extensive disease with at least one distant metastasis).

At midpoint of the study (Aug/2007), there were 50 patients with metastatic breast cancer, and 21 were live at the conclusion of the study in Dec/2010.

Chemotherapy is primary treatment for metastatic breast cancer and causes metastatic regression in just 1-3% of cases. Patients treated with our mushroom extracts experienced metastatic regression in 20% of cases.

Five-year survival rate in patients with metastatic breast cancer (measured from the first diagnosis), in the USA was at 14% (Lippman, Harrison’s Principles of Internal Medicine, 2005). Patients treated with intensive mycotherapy had a survival rate of 20% (measured from the start of mycotherapy!).

Unfortunately, the sample sizes were too small to reach several conclusions. However, it is evident that (1) taking large doses of medicinal mushroom extracts for a long time is completely safe, (2) the initial effects of starting mycotherapy are very pronounced (with at least using for 40-60 days), and (3) the effects are dose dependent, with larger doses/longer use yielding better results.

See the Presentation (given at the 6th International Mushroom Conference, Zagreb 2011)

 

Neven Jakopovich presents the results of the study at the 6th International Medicinal Mushroom Conference in Zagreb, 2011. The conference was hosted and organized by Myko San company.
Neven Jakopovich presents the results of the study at the 6th International Medicinal Mushroom Conference in Zagreb, 2011. The conference was hosted and organized by Myko San company.