NKCP :

NKCP, a purified filtrate of Bacillus subtilis var. natto culture is a food based extract of “natto”, a Japanese traditional fermented food made from soybean. Purification to remove most of distinctive odor of natto and its vitamin K2 yields an easy-to-eat food that has a wide variety of uses as a functional food. NKCP contains proteolytic enzymes secreted by Bacillus subtilis var. natto (Bacillus subtilis var. natto-produced protein), which balance clotting mechanisms in the blood. 

In vitro and clinical studies have demonstrated that the consistent intake of NKCP over a prolonged period helps to maintain normal circulation. The safety of NKCP has been demonstrated in safety studies. *In vitro (Latin for "within the glass") refers to the technique of performing a given procedure in a controlled environment outside of a living organism.

Patents (Production process for purified filtrate of Bacillus subtilis var. natto culture)
Japan (No.3532503)

NKCP stands for Purified Natto Culture Filtrate. NKCP tablets launched April 2001. Daiwan Health Development Inc. (DHD) began export of NKCP on November 2001.

Development Background

The “People's Health Promotion Campaign for the 21st Century (Healthy Japan 21)” was launched in 2000 by the Ministry of Health, Labour and Welfare. The purposes of this campaign are to reduce premature death, prolong optimal health, and improve the quality of life. Simply put, it focuses on longevity accompanied by optimal health. Cardiovascular and cerebrovascular diseases account for about 30% of causes of death in Japan, about 25% in the world. To lower mortality rates while in the prime of life, it is very important to prevent cardiac and cerebrovascular diseases. 

A patient with cardiovascular disease may evade a fatal cardiovascular event, but eventually their quality and duration of life will be compromised.

Contemporary lifestyles choices are associated with an increased risk of thrombus formation and subsequent ischemic heart disease and cerebrovascular disease. It has recently been demonstrated that the etiology of travel induced thrombosis, also known as “economy class syndrome”, is Deep Vein Thrombosis (DVT), which may result in a pulmonary embolism. DVT may develop as a result of prolonged sitting in cramped quarters related to both air and auto travel. Additionally, common ailments such as stiff shoulder muscles and leg edema may be caused by insufficient peripheral circulation, caused by excess blood viscosity. The key to preventing DVT and insufficient peripheral circulation is to balance coagulation and fibrinolysis, to prevent thrombus formation and to enhance overall cardiovascular health and longevity. NKCP may be a critical component to achieve such balance.

Development

NKCP was developed based on the traditional Japanese food, natto. Natto contains constituents that enhance the fibrinolysis system in favor of clot lysis. Naturally occurring proteolytic enzymes produced by Bacillus subtilis var. natto dissolve clots in a balanced manner, without causing excessive blood thinning. Recent research has shown that other additional constituents from Bacillus subtilis var. natto produce a substance that acts to inhibit blood coagulation, thereby complementing the known fibrinolytic activity to improve blood viscosity. Based on these insights, it is probable that consistent consumption of natto may lower the overall risk of thrombus formation. However, compliance of such a recommendation may be poor, due to natto's potentially objectionable odor and flavor. Additionally, natto consumption may be contraindicated for patients on anti-coagulant therapy due to its high vitamin K2 content. Furthermore, the proteolytic enzyme content may vary greatly in commercially available natto, resulting in inconsistent anti-coagulant activity.

NKCP was therefore developed to provide a raw material for food products which corrects the drawbacks of natto. NKCP is produced by fermenting the bacillus in a liquid medium containing soybean extract and then partially purifying the peptidase. The odor, bacterial body and vitamin K2 content are reduced to a negligible level. NKCP is designed to contain a constant amount of peptidase.

Characteristics of NKCP

•　NKCP is extracted from Bacillus subtilis var. natto and it is free of the undesirable odor and viscous texture of natto.
•　NKCP's purpose is three-fold.
To function as (1) an anticoagulant (2) thrombolytic and (3) decreases blood viscosity.
•　The majority of the vitamin K2 has been eliminated, therefore it is less antagonistic to other drugs such as warfarin.
•　NKCP is standardized to contain specific levels of proteolytic enzymes secreted by Bacillus subtilis var. natto.
•　The principal functional enzyme (protease) is stable at pH 6.0-10.0 and at temperatures 60℃ or below.
•　The safety of NKCP has been confirmed in many animal and human studies.
•　The production process for this purified filtrate of Bacillus subtilis var. natto culture is registered under Patent No. 3532503 in Japan.

Suggested daily dose


125-500mg/day


Mechanism of Action

NKCP is shown to have the following effects:
a. Inhibiting thrombus formation in vitro and in vivo. 
b. Decreasing the viscosity of blood in vitro and in vivo. 
c. Lysing thrombi in vitro and in vivo. 
*In vivo (of processes) performed or taking place in a living organism."fluid transport was measured in vivo"

The coagulation/fibrinolysis system is comprised of a series of complicated reactions designed to maintain the balance between healthy circulation and prevention of excess bleeding. Many factors can influence this system, however, it is not easily disrupted. In the event that the system shifts towards excess thrombus formation, it is challenging to return the system back to balance. Because it is difficult to lyse a formed thrombus, the emphasis should be placed on prevention of thrombus formation rather than on thrombolysis.

By inhibiting thrombus formation and decreasing blood viscosity, orally administered NKCP helps maintain balance, shifts blood away from clot formation, and enhances circulation throughout the body.



Scientific Data

1. NKCP Actions

Reduced risk of thrombus formation requires: 1) clot formation prevention 2) maintenance of normal blood viscosity and 3) lysing blood clots (thrombi).
NKCP, derived from Bacillus subtilis var. natto, has been shown to perform these three functions.

2. Supporting Scientific Data

(1) Anticoagulant effect

▶Anticoagulant effect of NKCP in human blood.
▶Anticoagulant effect of NKCP in rat model of thrombosis formation.

(2) Action of preventing increase in blood viscosity.

▶Effect of NKCP on the viscosity of human blood

(3) Thrombolytic effect.

▶Thrombolytic effect of NKCP on artificial thrombi.
▶In vivo thrombolytic effect of oral NKCP in experimental thrombolysis model.
▶Identification of proteases derived from Bacillus subtilis var. natto related to thrombolysis.

3. Clinical Study Results

▶Effect of Bacillus subtilis var. natto-derived protein on the human blood coagulation/fibrinolysis system.

Safety

Single-dose:
LD50＞5,000mg/kg

Repeated-dose:
NOAEL
Males: > 1,325mg/kg body weight/day.

Females: > 1,541mg/kg body weight/day.

Mutagenicity:
Negative (± metabolic activation).

Antigenicity (guinea pigs): Negative for active systemic anaphylactic reaction (ASA) and passive cutaneous anaphylactic reaction (PCA).

Effect on bleeding time (rats):
In rats orally given NKCP, a 0.5mm incision was made in the tail tip after 1 hour to measure bleeding time. NKCP at 300mg/kg did not prolong the bleeding time.

Interaction with warfarin (rats):
NKCP at 250mg/kg was administered into the duodenum by the in situ loop method in rats, in which bleeding time was delayed by treatment with warfarin, and blood collected after 6 hours was measured for coagulation time. The warfarin treatment significantly prolonged the coagulation time in comparison with the control group, but no added delay of coagulation was observed in the warfarin + NKCP treatment group compared with warfarin treatment group.

Long-term administration (humans):
Twenty-three healthy adults were given NKCP at 250mg/day for 12 weeks, and no clinically significant adverse events were observed. There were no statistically significant changes in hematological or biochemistry tests.

Five healthy adults were given NKCP at 750mg/day for 6 consecutive weeks to study and observe changes in laboratory test values (hematological tests, biochemistry tests, and blood coagulation/fibrinolysis parameters) and adverse events. As a result, ELT shortened, t-PA decreased, and thromboplastinogen activity test (TAT) increased but all values were within normal range. In addition, no adverse events were observed, suggesting NKCP safety.

High dose administration(humans):
Eight healthy adults were given NKCP at 1,250mg/day for 7 consecutive days. Observation of clinical signs and laboratory tests were utilized to evaluate NKCP safety. There were no clinically significant adverse events. There were no abnormal changes in hematological or biochemistry tests.

Assays

1. Peptidase Activity (synthetic substrate method)

The sample solution is warmed at 37°C with the synthetic chromogenic substrate S-2251 (H-D-valyl-L-leucyl-L-lysine-p-nitroanilide dihydrochloride) as a substrate and the absorbance at 405nm is determined. The enzyme activity is defined as 1 unit when 1nmol of p-nitroaniline per minute is released.
*An assay is an investigative (analytic) procedure in laboratory medicine, pharmacology, environmental biology and molecular biology for qualitatively assessing or quantitatively measuring the presence, amount, or functional activity of a target entity (the analyte).

2. Bacillus subtilis var. natto-produced Protein

ELISA (enzyme-linked immunosorbent assay) uses rabbit-specific antibodies to the Bacillus subtilis var. natto-produced protein, responsible for the peptidase activity, to measure the amount of antigen reacting with the specific antibodies.

▶Anticoagulant effect of NKCP in human blood

Thirty micro litters of the test substance, NKCP, was added to 3mL of venous blood collected from healthy volunteers who had given informed consent, followed by inversion for mixing. The mixture was warmed at 37°C for 250 seconds and centrifuged, and the supernatant was measured for fibrin monomers (FM) indicative of thrombus formation, using a latex immunity analyzer. The concentration of FM was 160 ± 29.3μg/mL after addition of the control physiological saline and 6.0 ± 1.1μg/mL after addition of heparin sodium (0.5IU/mL). After addition of NKCP at concentrations from 0.005mg/mL to 0.5mg/mL, the concentration of FM decreased dose-dependently, but plateaued at 0.05mg/mL.

The 54th Study Meeting of Rheology 2006;
Department of Legal Medicine, Dokkyo Medical University School of Medicine

▶Anticoagulant effect of NKCP in rat model of thrombosis formation

The anticoagulant effect of NKCP was studied in a rat model of thrombus formation. In this model, platelet aggregation was caused by injuring endothelial cells of the abdominal descending aorta to induce thrombus formation. NKCP was administered into the duodenum 2 hours after producing the thrombus formation model using in situ loop method. Blood was collected from the abdominal aorta 6 hours after administration to determine activated partial thromboplastin time (APTT) and prothrombin time (PT) as indicators of endogenous and exogenous coagulations, respectively. APTT was 33.5 ± 2.4 seconds for the control group (administered physiological saline), 52.0 ± 4.5 seconds for the NKCP 100mg/kg group, and 63.3 ± 2.9 seconds for the NKCP 250mg/kg group. A significant coagulation delay was observed in the NKCP treatment groups as compared to control. PT was 16.7 ± 0.5 seconds for the control group, 20.6 ± 0.9 seconds for the NKCP 100mg/kg group, and 21.3 ± 1.7 seconds for the NKCP 250mg/kg group. As with the APTT, a significant coagulation delay was observed in the NKCP group as compared to control. These results suggest NKCP inhibits thrombus formation.

Research and Development Department, Daiwa Pharmaceutical Co., Ltd., 2001

▶Effect of NKCP on the viscosity of human blood

Eight healthy male adults who had given informed consent were given a single dose of placebo or NKCP at 1,250mg, and blood was collected at various points over 240 minutes to measure changes in blood viscosity. The viscosity of blood was determined by Hitosugi et al.'s method using an oscillating viscometer.
In the NKCP group, there was a significant decrease in blood viscosity at 105 and 180 minutes as compared to baseline. There was a significant decrease in blood viscosity in the NKCP group as compared to placebo at 180 minutes.



Effect of NKCP on the viscosity of human blood:
Department of Legal Medicine, Dokkyo Medical University School of Medicine.

▶Thrombolytic effect of NKCP on artificial thrombi

A small amount of NKCP was added to test tubes containing an artificial thrombus in physiological saline. The thrombus began to lyse within several minutes and was almost completely lysed at 3 hours. To lyse is to cause dissolution or destruction of cells by lysins. Lysis is the disintegration of a cell by rupture of the cell wall or membrane.



Thrombolytic effect of NKCP on artificial thrombi:
J. Pharmacol Sci 2005; 99: 247-251.

▶In vivo thrombolytic effect of oral NKCP in experimental thrombolysis model

The thrombolytic effect of NKCP on rat model was observed in a 14 week study. The NKCP was evaluated in two groups, in a mix of 0.2% and 1% in feed, as compared to a control group. Thrombolysis was evaluated using a He-Ne laser induced thrombosis model in mesenteric microvessels. The size of the artificially produced thrombus was measured from the time of formation to evaluate the thrombolytic effect of NKCP.

Thrombolytic activity clearly increased dose-dependently in the NKCP treatment groups compared with the control group. There was an 82% decrease in thrombus volume in the control group, as compared to 67% decrease in the 0.2% NKCP group, and a 51% decrease (statistically significant) in the 1% NKCP group. The extent of thrombolysis in the 1% group was equivalent to that seen in animals treated with a bolus intravenous infusion of 0.2mg/kg of tissue plasminogen activator (t-PA). Based on the body weight and feed intake of rats used in the study, the dose of NKCP was calculated to be about 160mg/kg/day for the 0.2% NKCP feed and 800mg/kg/day for the 1% NKCP feed.



In vivo thrombolytic effect of oral NKCP in experimental thrombolysis model:
Pathophysiol Haemost Thromb 2003; 33: 138-143.

▶Identification of proteases derived from Bacillus subtilis var. natto related to thrombolysis

The thrombolytic effect of NKCP is attributable to a proteolytic enzyme named bacillopeptidase F, produced by Bacillus subtilis var. natto. Bacillopeptidase F is shown to be one of five proteases produced and secreted by Bacillus subtilis var. natto (Table 1).



Identification of proteases derived from Bacillus subtilis natto related to thrombolysis:
1)Journal of Bacteriology 1990; 172: 1019-1023
2)The Journal of Biological Chemistry 1990; 265: 6845-6850
3)Journal of Bacteriology 1990; 172: 1470-1477
4)Experientia 1987; 43: 1110-1111
5)The Journal of Biological Chemistry 2001; 276: 24690-24696
6)Mol Gen Genet 1990 May; 221(3): 486-490

▶Effect of Bacillus subtilis var. natto-derived protein on the human blood coagulation/fibrinolysis system

A total of 23 adults, including patients with metabolic diseases related to thrombosis, were given 250mg NKCP for two consecutive months. Coagulation/fibrinolysis parameters and symptoms were evaluated 1 and 2 months after starting treatment.

Euglobulin lysis time (ELT) significantly decreased from the baseline value at 1 and 2 months, and t-PA significantly increased at 2 months, showing the accelerated fibrinolysis system. However, the measured ELT and t-PA values were within the normal ranges. Fibrinogen degradation products (FDP) significantly decreased 1 month after starting ingestion but returned to the baseline value at 2 months.

Symptoms of shoulder stiffness significantly improved from baseline at 1 and 2 months.



Effect of Bacillus subtilis natto-derived protein on the human blood coagulation/fibrinolysis system:
Journal of the Japanese Society of Biorheology 2004; 18 (1).

NATTOKINASE

Other Names: BSP, Extrait de Natto, Fermented Soybeans, Haricots de Soja Fermentés, Natto de Soja, Natto Extract, Nattokinasa, NK, Soy Natto, Subtilisin NAT.

NATTOKINASE OVERVIEW INFORMATION
Nattokinase is an enzyme (a protein that speeds up biochemical reactions) that is extracted from a popular Japanese food called natto. Natto is boiled soybeans that have been fermented with a bacterium called Bacillus natto.

Natto has been used as a folk remedy for diseases of the heart and circulatory system (cardiovascular disease) for hundreds of years. Nattokinase, the chemical in natto that is probably responsible for its effects, was discovered by a University of Chicago researcher, Dr. Hiroyuki Sumi.

You won’t find nattokinase in soy foods other than natto, since nattokinase is produced through the specific fermentation process used to make natto.

Nattokinase is used for cardiovascular diseases including heart disease, high blood pressure, stroke, chest pain (angina), deep vein thrombosis (DVT), “hardening of the arteries” (atherosclerosis), hemorrhoids, varicose veins, poor circulation, and peripheral artery disease (PAD).

It is also used for pain, fibromyalgia, chronic fatigue syndrome, endometriosis, uterine fibroids, muscle spasms, infertility, cancer, and a vitamin-deficiency disease called beriberi.

How does it work?
Nattokinase decreases the ability of blood to clot. This "thins the blood" and might protect against conditions caused by blood clots such as stroke, heart attack, and others.

NATTOKINASE USES & EFFECTIVENESS 

Possibly Effective for:
High blood pressure. Research suggests that taking nattokinase (NSK II, Japan Bio Sciences Laboratories Company Ltd., Japan) daily for 8 weeks can reduce blood pressure in people with high blood pressure.

Insufficient Evidence for:
Deep vein thrombosis (DVT). There is some evidence that taking a specific combination product (Flite Tabs) might decrease the chance of getting a blood clot in the legs during long plane flights. This product combines a blend of 150 mg of nattokinase plus pycnogenol. Two capsules are taken 2 hours before the flight and then again 6 hours later.
Heart disease.
Stroke.
Angina.
“Hardening of the ateries” (atherosclerosis).
Hemorrhoids.
Poor circulation.
Varicose veins.
Peripheral artery disease (PAD).
Pain.
Fibromyalgia.
Chronic fatigue syndrome.
Endometriosis.
Uterine fibroids.
Muscle spasms.
Infertility.
Cancer.
Beriberi.
Other conditions.

More evidence is needed to rate the effectiveness of nattokinase for these uses.

NATTOKINASE SIDE EFFECTS & SAFETY
Nattokinase is LIKELY SAFE when taken by mouth in amounts commonly found in foods. Nattokinase is a natural component of the soy food natto. It has been routinely consumed in Japanese cultures for hundreds of years.

Nattokinase is POSSIBLY SAFE when taken by mouth as medicine. Taking two doses of a specific product containing nattokinase (Flite Tabs) seems to be safe. However, it is not known if taking more than two doses is safe.

Special Precautions & Warnings:
Pregnancy and breast-feeding: There is not enough reliable information about the safety of taking nattokinase if you are pregnant or breast-feeding. Stay on the safe side and avoid use.

Bleeding disorders: Nattokinase seems to act like a “clot thinner” and might make bleeding disorders worse. Use with caution.


Surgery: Nattokinase might increase the chance of bleeding too much during or after surgery. Stop taking it at least 2 weeks before a scheduled surgery.

NATTOKINASE INTERACTIONS 

Moderate Interaction: Be cautious with this combination
Medications that slow blood clotting (Anticoagulant / Antiplatelet drugs) interacts with NATTOKINASE.

Nattokinase can decrease blood clotting. Taking nattokinase along with medications that also slow clotting might increase the chances of bruising and bleeding.


Some medications that slow blood clotting include aspirin, clopidogrel (Plavix), diclofenac (Voltaren, Cataflam, others), ibuprofen (Advil, Motrin, others), naproxen (Anaprox, Naprosyn, others), dalteparin (Fragmin), enoxaparin (Lovenox), heparin, warfarin (Coumadin), and others.

NATTOKINASE DOSING

The appropriate dose of nattokinase depends on several factors such as the user's age, health, and several other conditions. At this time there is not enough scientific information to determine an appropriate range of doses for nattokinase. Keep in mind that natural products are not always necessarily safe and dosages can be important. Be sure to follow relevant directions on product labels and consult your pharmacist or physician or other healthcare professional before using.

User Reviews

1. Comment:
Have AFib with episodes maybe 1 in 6 months or longer. I've been taking "Healthy Origins" Nattokinase with Vit K removed 100mg 2x's day. I feel that my circulation has improved and I also have T2 diabetes and worry a lot about it and believe it's amazing. I also don't worry about not taking pradaxa or any other blood thinner ( to prevent strokes and heart attacks) that could be harmful and dangerous, i trust Nattokinase! Healthy Origins brand has the ingredients I want and doesn't what I don't want! 

2. Comment:
Seven years ago I had a heart catherization completed.I was diagnosed with a 50% blockage of the LAD (The widow maker) Three and one half years ago I had a 64 bit CAT Scan of the Heart, this also revealed a 50% blockage of the LAD. The interesting part is that approx. 6 years ago on the recommendation of my son who incidentally has his PharmD I began a regiment of NattoZyme 100mg 2000FU taking 1 capsule twice daily. now seven years later the blockage has not progressed at all. Strange / Coincidental / Fact. I do not know but it worked for me. i am a believer!

3. Comment: 
Why haven't further studies been done to provide more data for determining effectiveness of this supplement?

4. Comment:
Suffered a brain stem stroke 3/2012. After I slowly got off Pradaxa, a prescription anti-coagulant, I thought I needed something other than low dose aspirin, so I tried Nattokinase. No side effects, but I noticed shaving & bleeding cuts took a few seconds longer to coagulate. I switched to a plant base diet, low dose vitamin K2 & magnesium glycinate. No 2nd stroke nor TIAs to report since 3/2012.

5. Comment: 
Nattokinase is not only a folk remedy for stroke but also is currently a beneficial treatment for ischemic stroke esp. TIA.

6. Comment:
Update from my post of 6/2/12. I now have been taking natto for 7 years for my antiphospholipid syndrome and it continues to work very effectively for me, without any side effects nor further stroke/tia. I have had close calls, a few perhaps mini-tia's, but as said previously, compared to having monthly tia's, I can deal with this. As it still is not approved by the FDA (I so wish they would do testing for it), my doctors take it casually. One doctor would not see me, another does accept it. However, I know it helps. One note of caution: Do be very careful with your vitamin K intake because natto will not be absorbed if too much is taken in. All dark, leafy vegetables. I was taking a protein supplement which did not state it contained high amounts of vitamin K and that is when I had a close call. Also, do be sure you are buying high grade natto and with the vitamin K removed. The top grade ones can be bought through the internet vitamin stores, I use NutriCology. (hope that's ok to post here). Good luck!

7. Comment: 
works well as a blood thinner.

8. Comment: 
Nattokinase is a natural supplement that has worked well for me. There has been so much research for Nattokinase especially in Japan. Check out this video for more information. https://www.youtube.com/watch?v=TYp EKGd7iIQ


9. Comment:
Have had consistant atrial fib for over 3 years now and have chosen to take Nattokinase rather than prescription blood thinners, after reading about this alternative. Of course the Docs (cardiologists) dismiss it and not one has even heard of or acknol'd it. In any case, I've taken 2 of the 2,000 unit each early AM (for an empty stomach) since diagnosed and have had no problems. Recent cardiac cath shows NO blockages, clean arteries/veins. Have had none of the typical swelling of ankles, nor other symptoms. Not sure if I'm taking enough to prevent that everpresent threat of stroke from AF, but so far so good. I am Stuart at DrinksScotch@gmail.com

10. Comment:
Decided to use Nattokinase when side effects from Plavix became intolerable. While using it, had no TIAs though I did with Plavix. Stopped the natto' briefly to try to return to Plavix due to Dr's wishes, had visual disturbances and tinnitus immediately. Also all my joint and muscle pain mysteriously goes away on the Nattokinase, some skin tags disappeared, and I just feel better. I take one Doctors' Best 2000 mg every other day just before sleep or in middle of the night to assure no food interferes with its absorbtion. Even if it doesn't replace Plavix, I will take it, as it erradicates my 24/7 knee/body inflamation pain totally. 

11. Comment: 
I have been taking this for help with menstrual cramps. I find it to be effective, but it is hard to make sure to take it on an empty stomach.

12. Comment: 
had a mini stoke prescribed wafferin and asprin I have canceled them and have been taking natto for five years now feel good.

13. Comment:
Condition: Protein-S Deficiency My husband was told he couldn't have the IV antibiotic treatment he needed for his neurological lyme disease because he was so severely deficient in anti-clotting factor, protein-s. Normal range: 60-145, His result: 9! They said they'd retest in a month. I read everything I could find on protein-s and decided to give him nattokinase for it's high vitamin k content. When they retested him a month later, his level had risen from a 9 to a 95, well within normal range and he was able to go on IV antibiotics for his lyme disease, which has given him his life back.

14. Comment: 
Heart has remained in Atrial Fibrillation for 9 years. Took Coumadin following several orthopedic surgeries, but am not willing to take it long term. Have taken Nattokinase for at least 3 years. Recently had a cardiac catheterization and they found no blockages. I attribute that to the nattokinase.

15. Comment: 
Have AF. No blood clots in over 10 years! No side effects.

16. Comment: 
I have atrial fibrillation. Nattokinase is my treatment of choice, as opposed to coumadin or xarelto. NO SIDE EFFECTS.

17. Comment:
I have Peripheral Artery Disease. I got it in my late 40s due to blood coagulation problem. I also had very high cholesterol (over 300+ my entire life). I had difficulty walking by age 50. I put myself on high dose nattokinase and serrapeptase and the symptoms disappeared within 6 weeks. You will know it is working within 1 month. I eat as much cholesterol as I want. I have been on this over 6 years. I am on maintenance dose. Just experiment with dosage over time to get the minimum dose and cost. Please be aware that conventional medicine perpetuates lies about cholesterol. Just take the time to look at the studies. If the arteries are clean enough, you won't have heart disease. Ignore the cholesterol propaganda. They have Nattokinase made from bacteria and fungus, with and without vitamin K. Both work but I prefer the Nattokinase with Vitamin K. Without vitamin K is for people on prescriptions drugs. I take Best Nattokinase and Serrapeptase. I buy from Swanson vitamin because they are cheap. iI'm amazed they don't have Heart disease category for this. Chinese have done thousands of studies on effectiveness of fibrolytic enzymes for heart attack and stroke prevention. 

18. Comment:
I've suffered high blood pressure and high triglyceride levels. I have suffered what it used to a "minor" heart attack in the past due to a blockage of artery in my heart. But since taking nattokinase, I have been able to control my blood pressure without drug prescriptions, and my triglycerides and the good cholesteral level are normal. And all this, without going to extremes in watching my diet.

19. Comment:
I was diagnosed with antiphospholipid syndrome along with low factor VIII levels in December of 2007. I had had numerous tia's previously. Because of the low factor VIII levels I was not a candidate for wayfarin but my hematologist put me on coumadin. After 4 months it was determined the coumadin was also imcompatible; my hematologist literally told me to get my will in order as nothing could be done. I found out about nattokinase and found a pharmaceutical grade brand containing no vitamin K. Have been taking 400 mg. daily, divided, without food, for 4 years now. No bad side effects at all. Cannot take aspirin with it. The good benefits? I have had a few tia's, yes - perhaps 5 total in 4 1/2 years. But when you compare that to the average of perhaps 4 per month was having in the 6 months prior to being diagnosed, this is amazing. It took the doctors that long to diagnose me and I was so fortunate not to have had a major stroke. I also watch my diet, not eating vegetables high in vitamin K. Other than that I feel I owe nattokinase my life. I so wish it would be properly tested and therefore approved.

20. Comment: 
I have MTHFR C667T and my husband has MTHFR A1298C and we miscarried 4 babies on Lovenox. So I tried nattokinase 2 times a day and now have a beautiful princess! I still take it to prevents clots and fibroids.

21. Comment: 
I am using nattokinase and serrapeptase to shrink some large uterine fibroids. So far it is working, I estimate that since my fibroids are large it will take up to 6 months for my belly to flatten. but so far i can feel they are getting smaller.

22. Comment:
I had a six inch clot in my inferior vena cava which blocked 90 percent of the blood flow. My hematologist stated after examining me that he could do nothing and that I was a walking time bomb. I react badly to Coumadin and I had DVT in my right leg which blocked blood flow on all outside veins from ankle to groin so I chose a Nattokinaise that did not contain vitamin "K". The major clot is almost totally dissolved and the blocked veins in my leg have regained much of their blood flow. I have been taking 300 mg. twelve hours apart. I have not experienced the headaches I had with Coumadin nor the elevation in blood pressure in excess of 240 systolic, which happened on several occasions. I was careful not to take anything that might act as a blood thinner and not to consume green vegetables rich in vitamin "K" during my use of Nattokinaise. Now that the clot has been greatly reduced, my doctor placed a "Greenfield" filter in the vena cava, above the blood flow from the right leg; I shall worry less about the remaining clots. I thank GOD for the people that make this type of Nattokinaise as I was concerned that continued use of Coumadin presented a stroke possibility for me. 

23. Comment: 
great energy , sever heart palpitations.

24.  Comment:
Important to take on empty stomach or between meals, otherwise it will become a digestive aide. I am using for diabetic restless legs and results were almost instantaneous.Developed surface clots at ankles so doubled dose on Dr's advice.Working good! Using Dr's Best Nattokinase,( 2000FU caps.)- 2 capsules twice a day. Added bonus is reduction of blood pressure allowing me to cut medicine in half and possibly stoping it soon. My memory is much much improved also.I am 71 and I'm sure the improved circulation is good for the whole body. To me, Nattokinase is like a miracle.

25. Comment:
I was tested positive for the prothrombin gene mutation and MTHFR. I got pregnant twice and miscarried and was told the mutation could be causing it. I was told that I should start heparin and try to get pregnant. I started researching and found nattokinase and thought I would try that before heparin. I started taking 2 a day in January of 2008 and February 2008 got pregnant. Continued taking Nattokinase through my pregnancy without taking heparin. I have since had two healthy girls. My high risk doctor told me on many occasions that I'm taking a chance with my life but I really felt that he just had no knowledge of nattokinase. I am still taking 1 tablet a day now to prevent future clots and I feel great.

26. Comment: 
77 year old woman -Used for atherosclerosis. Told that my carotid and several other arteries were 90+% blocked and was scheduled for arteriogram 2 months later. Started Nattokinase 4 times/day. Vascular surgeon says only 20-25% blocked when arteriogram happened. Amazing.

27. Comment: 
have tried everythng for hypertension and blood press still out of control. took my bp monday 151 over 101. started on natto 100mg once a day, third day bp 121/76 wow! fourth and fifth day 121/82, 126/76. it works! i'm 36 years old and have had high bp for 8 years, wish i would've found out about natto sooner!

Frequently Asked Questions
Answers to frequently asked questions on natto NKCP

Who first isolated the active component of natto?

The main active component of the food natto is an enzyme that was first isolated and identified by Dr. Hiroyuki Sumi in 1980, a researcher at the Japan Ministry of Education. Dr Sumi named this enzyme "nattokinase", commenting at the time that it showed "a potency matched by no other enzyme." NKCP is a high quality proprietary extract of natto that contains a high concentration of nattokinase.

Is NKCP a medicine?

NKCP has no direct effect on blood viscosity. What it actually does is help increase the efficiency of the body's own control mechanism. By doing this, if blood viscosity is too high, the body is able to naturally reduce it. It does not interfere with normal blood coagulation.

Why use NKCP for maintaining optimum blood flow when I can use aspirin which is much cheaper?

Aspirin is an over-the-counter medicine that has a direct effect on blood coagulation. Affecting body physiology directly like that can be problematic. For example, although the there are health benefits from aspirin use (and you must follow your doctor's advice) it should not be forgotten that aspirin is a strong medicine and that, although it is overall a relatively safe medicine considering how much has been taken around the world, long-term use has been linked to pancreatic cancer, gastrointestinal irritation, stomach bleeding, stomach ulcers (caused by bacteria), allergic hypersensitivity in asthmatics, brain haemorrhage and inflammation (Reye's syndrome). NKCP is not a medicine and does not have a medicinal action; what it does is helps the body to control its own blood viscosity. This is why it does not affect blood coagulation and can safely be taken, even long-term, without increasing the risk of haemorrhage. It can also be taken in conjunction with anti-coagulants such as Warfarin, although you are always advised to clear any food supplement with your doctor if you are on medication or have a specific medical condition.

How safe is NKCP to take?

NKCP is an extremely safe natural health supplement with no reported contra-indications or side-effects, even in large amounts over long periods of time. If you are on medication, however, you should always check with your doctor before adding any food supplement to your diet.

If I am taking NKCP to help with circulation during a long haul plane flight, how soon before should I start taking it?

NKCP needs to be taken at least 8 hours before a flight for it to be effective. Ideally, a person should start taking a daily dose a few days before.

Can NKCP help those with high blood pressure?

There are many reasons for high blood pressure and these should be explored with your doctor. That said, Dr. Maeda at Daiwa Pharmaceutical has stated that improved blood flow in peripheral vascular regions could help lower high blood pressure. In fact, in a Japanese NKCP clinical trail on humans, one subject with high blood pressure had it lowered. So it is certainly worth a try but also check with your doctor.

Can NKCP help patients with carotid artery stenosis?

Carotid artery stenosis is a disease causing atherosclerosis in the carotid artery, in which　the carotid artery becomes narrower. Progressive hardening of the arteries may result in the clot formation in the artery. These clots sometimes cause stroke or myocardial infarction.　We have no clinical data about the effect of NKCP on the patients with carotid arterystenosis. However, NKCP has been shown to be effective in inhibiting thrombus formation (prolongation of APTT and PT) and in promoting thrombolysis in animal experiments. Also, a clinical study reported that NKCP enhanced fibrinolytic activity by reducing the Euglobulin Lysis time (ELT). In addition, several studies reported NKCP improved stiff shoulders and other symptoms by improving the blood flow, and lowered the blood pressure. Based on these studies, we recommend the patient take NKCP, since this product is considered to be beneficial in carotid artery stenosis.

Is NKCP suitable for vegetarians?

NKCP is vegetarian although, because it contains shellac wax as a tablet coating, it is not be suitable for vegans. (Shellac is classified as vegetarian by The Vegetarian Society in the United Kingdom.)

How should NKCP be taken?

It is recommended to take 2 to 4 x 125mg tablets once a day after the evening meal.

Why is this product tested on animals?


It is always regrettable to cause suffering, but it must not be forgotten that it is a legal requirement for new food extracts such as this one to be exhaustively tested, and that includes animals studies. All food supplements components and ingredients on the market today have at one time or another been required to go through this sort of testing, although if such testing was undertaken far enough back in time, or if a new product is merely a recombination of established tested ingredients, then a particular brand can state that it has not been tested on animals. Being an innovative food extract, this testing is highlighted with NKCP.

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NKCP® Nattokinase
Purified Filtrate of Bacillus Subtilis Natto Culture - the safe blood flow supplement

NATTO is a traditional Japanese food of fermented soybeans that has been eaten for many centuries. It is believed to be a health-giving delicacy and has an "acquired" taste and smell. New scientific research has discovered why natto is so good for health — the Bacillus subtilis natto (the good bacteria that form during the fermentation process) produce functional proteins (including nattokinase) which naturally and safely help the body to maintain optimum blood viscosity. Natto is, therefore, an important food for those wishing to help reduce the risk of thrombosis (blood clots) — a central factor in heart disease and strokes, two of the biggest killers in Western society today, and Deep Vein Thrombosis (DVT), a concern for those taking long haul airplane journeys. This is of particular interest for those wanting a safe, non-drug, Aspirin alternative for healthy blood flow.

The problem with eating natto is fourfold: the taste, smell and texture is unpleasant; it can contain large amounts of Vitamin K2 which interferes with medicines such as warfarin; activity can vary greatly from brand to brand; and this food is not widely available outside Japan. But now Daiwa Pharmaceutical has produced natto NKCP nattokinase — a standardized extract of natto bacillus culture (Bacillopeptidase F) in small odorless tablets with the vitamin K2 largely removed — so now everyone can enjoy the benefits of natto!

Summary
Summary information on natto NKCP.

Description

NKCP Nattokinase is a standardized extract of Bacillus Subtilis Natto Culture, a good bacteria found in the traditional Japanese delicacy called natto — fermented soybeans. The vitamin K2 content is also largely removed so that NKCP is compatible with anticoagulant medications such as Warfarin. NKCP tablets are odourless, easy to swallow, and contain no genetically modified ingredients.

Development

NKCP Nattokinase was developed and is manufactured by Daiwa Pharmaceutical Co. Ltd. in Japan, a small and innovative pharmaceutical company dedicated to producing natural food extracts and supplements for health and well being. Daiwa Pharmaceutical is renowned for being the developer and manufacturer of immune system supplement BioBran MGN-3.

Function

There a a number of enzymes that have been identified by Daiwa Pharmaceutical in NKCP natto extract, including Nattokinase, which have been shown to inactivate an unhealthy enzyme in the blood called plasminogen activator inhibitor (PAI-1), which increases as we get older. By helping to reduce PAI-1, NKCP greatly increases the efficiency of our body's natural control mechanisms for blood viscosity. The breakthrough with NKCP is that it has no medicinal effect — it does not directly act upon blood viscosity and does not directly dissolve blood clots. Rather, NKCP merely increases the efficiency of the body's own blood-control mechanisms. This means that this natural food supplement does not adversely affect blood coagulation — something that can happen using medicinal blood thinners such as aspirin.

Safety

NKCP Nattokinase safety has been confirmed by extensive testing, including independent safety examinations concerning acute toxicity, sub acute toxicity, mutagenicity, antigenicity and overdose. However, as with any food supplement, if you have a specific medical condition or are taking medication (especially relating to thrombosis or blood circulation), you are advised to consult with your doctor before taking NKCP Nattokinase.

Standard Dose


Base on evaluation of effectiveness and overdose tests on human, 250-500mg of NKCP (2-4 tablets) is recommended as the standard dose per day, taken after the evening meal.

Development
Development of Natto NKCP

Background

Heart disease and cerebrovascular disease (strokes) are respectively the first and the third leading causes of all deaths in developed parts of the world, with heart disease accounting for as much as 30% of all deaths and cerebrovascular disease accounting for nearly 10%. And these figures are only the tip of the iceberg in regards to the devastation of these diseases: for every one who actually dies from them, there are several others suffering from their ill effects — a stroke or a heart attack, for example, can leave a person bedridden. More mild symptoms might include neck stiffness, chronic headache, cold hands and feet, breathlessness, chest pains, poor mental function and dizziness.

Any focus on increasing length and quality of life must look, therefore, at methods to maintain a healthy vascular system and blood flow. This is fundamental to good health and was the reason why Daiwa Pharmaceutical Co. Ltd. started research into the health properties of a traditional Japanese food called natto which was supposed to be so beneficial to health and the circulation. What they discovered was that there is a good good bacteria in this traditional fermented food, B. subtilis natto, which contains enzymes that help the body to maintain optimum blood flow and viscosity by breaking down a substance in the blood called PAI-1 that interferes with the body's natural processes. PAI-1 concentrations increase as we get older which is why our blood gets more sticky as we age and why we are increasingly at risk from vascular diseases.

The problem with natto is is that it is a very unpleasant food to eat (it has an "acquired" taste/smell and a slimy texture); it contains high amounts of vitamin K2 which can interfere with anticoagulants such as Warfarin; the active enzyme content of natto can vary enormously from brand to brand (see below) and it is not easy to get hold of in Western countries. Daiwa Pharaceutical Co. Ltd., therefore, decided to extract the active ingredient (in the same way that good bacteria is extracted from yogart), remove the vitmain K2 in the process and to produce a standardized small tablet that would allow people to enjoy the benefits of natto without any of the drawbacks associated with it. They have called this "natto in a tablet" NKCP.

Comparison of good bacteria activity in 89 different natto brands found in Japan showing the variation in peptidase activity. Daiwa Pharmaceutical therefore decided to produced a standardized "natto in a tablet" equivalent.


Daiwa Pharmaceutical Co. Ltd.

Daiwa Pharmaceutical Co. is a new breed of pharmaceutical company focused entirely on the development of natural food solutions. Already renowned for producing what is now the leading serious immune system supplement, BioBran MGN-3, a food extract made entirely from rice bran, Daiwa Pharmaceutical have now developed the highest quality and most active Natto extract (sometimes called nattokinase, although this is just one of its components) — NKCP. What sets Daiwa Pharmaceutical apart from many other developers of innovative foods and food supplements is its dedication to high quality research and production. Daiwa is not afraid to developing natural (and therefore not patentable) products, something which most pharmaceutical companies would be reluctant to do.


Manufacturing Process

NKCP is manufactured by culturing B. subtilis natto -- the good bacteria isolated from the natto food -- in a liquid medium under conditions suitable for the production of the active protein. The cell bodies, insoluble matter and low-molecular weight substances (which include the characterisitic odour molecules) are then removed. The peptide concentration is then standardized and the NKCP is powdered and then pressed into tablets. Below is the manufacturing flow chart for NKCP.
NKCP Manufacturing Flow Chart
   
 
Cultivation of a particular strain of Bacillus subtilis natto culture on a medium of soybean protein, glucose and soluble starch

   
 
Removal of insoluble fraction by continuous centrifugation and filtration by celite

   
 
Removal of ultra-low-molecular weight fraction by ultrafiltration

   
 
Removal of cells by membrane filtration

   
 
Addition of excipients

   
 
Freeze-drying

   
 
Powdering & Tableting

Development

Development of Natto NKCP

1. Background Heart disease and cerebrovascular 
2. disease (strokes) are respectively the first and the 
3. third leading causes of all deaths in developed 
4. parts of the world, with heart disease accounting 
5. for as much as 30% of all deaths and 
6. cerebrovascular disease accounting for nearly 
7. 10%. And these figures are only the tip of the 
8. iceberg in regards to the devastation of these 
9. diseases: for every one who actually dies from 
10. them, there are several others suffering from their 
11. ill effects — a stroke or a heart attack, for example, 
12. can leave a person bedridden. More mild symptoms might include neck stiffness, chronic headache, 
13. cold hands and feet, breathlessness, chest pains, 
14. poor mental function and dizziness.
15. 
    
16. Any focus on increasing length and quality of life 
17. must look, therefore, at methods to maintain a 
18. healthy vascular system and blood flow. This is 
19. fundamental to good health and was the reason 
20. why Daiwa Pharmaceutical Co. Ltd. started 
21. research into the health properties of a traditional 
22. Japanese food called natto which was supposed 
23. to be so beneficial to health and the circulation. 
24. What they discovered was that there is a good 
25. bacteria in this traditional fermented food, B. 
26. subtilis natto, which contains enzymes that help 
27. the body to maintain optimum blood flow and 
28. viscosity by breaking down a substance in the 
29. blood called PAI-1 that interferes with the body's 
30. natural processes. PAI-1 concentrations increase 
31. as we get older which is why our blood gets more 
32. sticky as we age and why we are increasingly at
33.  risk from vascular diseases.
34. 
    The problem with natto is is that it is a very 
35. unpleasant food to eat (it has an "acquired" taste/
36. smell and a slimy texture); it contains high 
37. amounts of vitamin K2 which can interfere with 
38. anticoagulants such as Warfarin; the active 
39. enzyme content of natto can vary enormously 
40. from brand to brand (see below) and it is not easy 
41. to get hold of in Western countries. Daiwa 
42. Pharaceutical Co. Ltd., therefore, decided to 
43. extract the active ingredient (in the same way that 
44. good bacteria is extracted from yogart), remove 
45. the vitmain K2 in the process and to produce a 
46. standardized small tablet that would allow people to 
47. enjoy the benefits of natto without any of the 
48. drawbacks associated with it. They have called this 
49. "natto in a tablet" NKCP.
    
    Comparison of good bacteria activity in 89 different 
50. natto brands found in Japan showing the variation 
51. in peptidase activity. Daiwa Pharmaceutical there
52. fore decided to produced a standardized "natto in 
53. a tablet" equivalent.
54. Daiwa Pharmaceutical Co. Ltd.Daiwa Pharmaceutical Co. is a new breed of pharmaceutical company focused entirely on the development of natural food solutions. Already renowned for producing what is now the leading serious immune system supplement, BioBran MGN-3, a food extract made entirely from rice bran, Daiwa Pharmaceutical have now developed the highest quality and most active Natto extract (sometimes called nattokinase, although this is just one of its components) — NKCP. What sets Daiwa Pharmaceutical apart from many other developers of innovative foods and food supplements is its dedication to high quality research and production. Daiwa is not afraid to developing natural (and therefore not patentable) products, something which most pharmaceutical companies would be reluctant to do.
55. Manufacturing ProcessNKCP is manufactured by culturing B. subtilis natto -- the good bacteria isolated from the natto 
56. food -- in a liquid medium under conditions 
57. suitable for the production of the active protein. 
58. The cell bodies, insoluble matter and low-molecular 
59. weight substances (which include the characterisitic 
60. odour molecules) are then removed. The peptide 
61. concentration is then standardized and the NKCP 
62. is powdered and then pressed into tablets. Below 
63. is the manufacturing flow chart for NKCP.

NKCP Manufacturing Flow Chart
	Cultivation of a particular strain  of Bacillus subtilis natto culture  on a medium of soybean protein, glucose and soluble starch
	Removal of insoluble fraction  by continuous centrifugation  and filtration by celite
	Removal of ultra-low- molecular weight fraction  by ultrafiltration
	Removal of cells by  membrane filtration
	Addition of excipients
	Freeze-drying
	Powdering &  Tableting

Function

Biological function of Natto NKCP

1. Functional Ingredients

The functional ingredient in the food natto and the NKCP natto extract responsible for reducing the risk of thrombosis is a protein (sometimes called Nattokinase) produced by natto bacillus (B. subtilis natto). This is one of a number of proteins with differing molecular weights contained in natto. Its activity is confirmed through its ability to hydrolyze synthetic substrates for plasmin and fibrin. Improved thrombolytic activity has been demonstrated in a study on dogs given natto or partially purified extract of natto and in a study on humans taking natto.

NKCP has stable peptidase activity and its major active ingredient is a 34,000-45,000-dalton protein produced by the natto bacillus, belonging to the bacillopeptidase F family. The activity is stable at pH 6.0 ~ pH 9.0 under 60ºC and the optimum is pH 9.0. Its activity is measured by determining the ability to hydrolyze synthetic substrate for plasmin (S-2251) and its quantitative determination is carried out by measuring the amount of antigen reacting with the antibody specific for the protein, using ELISA.

Enzymes synthesized and excreted outside the cells by Bacillus subtilis after exponential multiplication.

Types	Gene	Properties
Subtilisin (alkaline) protease	apr	Molecular weight by SDS-PAGE: 20 kDa (1), 28 kDa (2) Nattokinase. Its casein decomposing activity and direct fibrinolytic activity have been confirmed. The ability to decompose and inactivate plasminogen activator inhibitor type 1 (PAI-1) has also been reported (2).
Neutral protease	npr	Major exoproteinase as with apr.
Extracellular protease	epr	Molecular weight by SDS-PAGE: 40-34 kDa
Metallo protease	mpr	Molecular weight by SDS-PAGE: 28 kDa (3)
Bacillopeptidase F	bpr	Molecular weight by SDS-PAGE: 47 kDa (3), 48 kDa (4). Secreted as a 92 kDa protein and converted into 80 kDa and 48 kDa proteins (4). Has high esterase activity as well as proteinase activity (5).

(1) Sumi, et al., Experientia, Vol. 43, 1110-1111, 1987.
(2) The Journal of Biological Chemistry, Vol. 276, pp. 24690-24696, 2001.
     Mol Gen Genet 1990 May; 221 (3): 486-90
(3) Journal of Bacteriology, Vol. 172, pp. 1019-1023, 1990.
(4) The Journal of Biological Chemistry, Vol. 265, pp. 6845-6850, 1990.
(5) Journal of Bacteriology, Vol. 172, pp. 1470-1477, 1990.

2. Mode of Action

NKCP has a thrombolytic effect in vitro and in vivo by ingestion and the effect is milder compared to the plasminogen activator (t-PA). NKCP has been confirmed to decompose and inactivate plasminogen activator inhibitor (PAI-1) in cell culture systems and it also reduces blood viscosity. Ingestion of NKCP reduces the plasminogen activator inhibitor (PAI-1) and helps the plasminogen activator (t-PA) to work efficiently. t-PA activates plasmin and reduces blood fibrin. Thus, NKCP facilitates the activation of fibrinolysis cascade reactions by reducing PAI-1 and helps maintain the balance between coagulation and fibrinolysis. Consequently, NKCP ingestion is unlikely to dissolve fibrin excessively.

Because of decreased PAI-1, NKCP fully activates the fibrinolytic cascade process and helps in maintaining the balance between coagulation and fibrinolysis of the blood. In other words, NKCP ingestion is unlikely to cause unexpected fibrinolysis. NKCP also reduces blood viscosity in vitro and in vivo. Although the association between this action and increased fibrinolytic activity is unknown, reduced blood viscosity is likely to contribute to maintaining sound blood conditions since it prevents stagnant blood flow.

3. Scientific Data on NKCP Functionality

a) Functionality in animals

i) Thrombolytic action of NKCP in rat thrombosis models

A comparative study was conducted by the Material Research Center to evaluate the thrombolytic action of NKCP by the in situ loop method using thrombosis models.

Thrombosis was produced in rats through platelet adhesion and agglutination, induced by injuries in the endothelial cells of the abdominal descending aorta. Six hours after the induction of thrombosis, the activated partial thromboplastin time (APTT) and prothrombin time (PT) were measured as indicators for blood clotting activity.

The APTT values were 33.5 ± 2.4 sec, 52.0 ± 4.5 sec and 63.3 ± 2.9 sec for the control, NKCP 100 mg/kg group and NKCP 250 mg/kg group, respectively, indicating a significant prolongation for NKCP. The PT values were 16.7?0.5 sec, 20.6 ± 0.9 sec and 21.3 ± 1.7 sec for the control, NKCP 100 mg/kg group and NKCP 250 mg/kg group, respectively, indicating a similar result for APTT. Since NKCP showed a significant prolongation of the coagulation time, as indicated above, its possible role in reducing thrombosis was suggested.

ii) In vivo effect of oral NKCP in experimental thrombolysis models
Laboratory of Physiology, Faculty of Nutrition, Kobe Gakuin University

The thrombolytic activity of NKCP after 14 weeks ingestion of feed containing 0.2% NKCP was evaluated in rat experimental thrombolysis models for arterial thrombosis mainly consisting of platelets using mesenteric microvessels.

The NKCP group showed an obvious increase in endogenous thrombolytic activity in a dose-dependent manner compared to the control group. The activity was equivalent to 0.2 mg/kg of tissue-type plasminogen activator (t-PA).

b) Activity in humans

i) Relation between NKCP ingestion and activity in humans
Daiwa Pharmaceutical Report

The effectiveness and safety of NKCP in a dose range between 1 and 8 tablets daily (125-1,000 mg on the NKCP basis) was evaluated in 40 healthy adults. The subjects ingested NKCP for a period from 1 day to 3 weeks. Various tests were performed to determine the optimum intake recommendations using the euglobulin lysis time (ELT) as the main indicator.

Doses of 2 tablets or more (250 mg in terms of NKCP) daily for a minimum of 4 days showed stable activity.

ii) Effect of the natto bacillus culture filtrate "NKCP" on the coagulative/fibrinolytic system: published as "Fibrinolytic and anti-thrombotic effect of the protein from Bacillus subtilis (natto) by the oral administration" - Japanese Society of Biorheology

The effect on the coagulation/fibrinolysis system of NKCP at a dose of 2 tablets daily (250 mg on NKCP basis) for 2 weeks was evaluated in 28 adults including patients with metabolic disease carrying the risk of thrombosis.

NKCP decreased the ELT by 10.1% without affecting the tissue-type plasminogen activator (t-PA) or the activated partial thromboplastin time (APTT), and this suggests that NKCP enhances fibrinolysis without intermediation by plasmin.

The effect of chronic use of NKCP at the dose of 2 tablets daily (250 mg on the NKCP basis) was evaluated in 23 subjects for 2 months. The ELT showed a significant decrease at 1 and 2 months and t-PA showed a significant increase at 2 months. Improvement in neck stiffness was observed at 1 and 2 months.

Changes in subjective symptoms due to NKCP intake

Symptom

Conditions

Before intake

At 1 month

At 2 months

Headache	Severe	1	1	1
	Moderate	7	3	4
	No symptom (including mild neck stiffness)	15	16	16
	Remarkable improvement	-	3	2
	Shirley-Williams multiple test	-	N.S.	N.S.

Neck stiffness	Severe	5	1	1
	Moderate	10	9	10
	No symptom (including mild neck stiffness)	8	9	11
	Remarkable improvement	-	4	1
	Shirley-Williams multiple test	-	P<0 .05="" td="">	P<0 .05="" td="">

Dizziness	Severe	0	0	0
	Moderate	6	4	4
	No symptom (including mild neck stiffness)	17	18	18
	Remarkable improvement	-	1	1
	Shirley-Williams multiple test	-	N.S.	N.S.

Figures represent the numbers of patients
Significant difference by Multiple Range Test: N.S. Not Significant, p<0 .05="" danger="" difference="" of="" rate="" significant="" span="" the="">

Changes in fibrinolysis/coagulation parameters due to NKCP intake (n=23)

Parameters	Normal values	Before intake	At 1 month	At 2 months
ELT(1)	6-12 hrs.	9.0 ± 1.3	8.1 ± 1.5**	8.0 ± 1.5**
t-PA(2)	≤ 10ng/mL	5.4 ± 2.6	5.8 ± 2.8	6.4 ± 2.2*
FDP(3)	≤ 4μg/mL	3.0 ± 0.7	2.0 ± 0.6*	3.0 ± 0.7

Figure represents ± Standard deviation. According to the significant difference by Duncan's Multiple Range Test, * shows the significant difference in 5 % of danger rate, and ** shows the significant difference in 1 % of danger rate.
1) Upper limit of measurement (ULM) is 12 hrs.
2) Lower limit of measurement (LLM) is 1.5 ng/mL.
3) LLM is 2 μg/mL.

iii) Anticoagulative/fibrinolytic effect of new natto extract
The 26th meeting of the Japanese Society of Biorheology

Natto culture filtrate without B. subtilis natto cells or vitamin K (hereafter abbreviated as NKCP) was subjected to gel filtration using a carrier such as Toyopearl HW-40F to extract NKCP with a molecular weight of 45 KDa. NKCP was shown to hydrolyze S-2251, a synthetic substrate specific to plasmin at 10 IU/mg/min. A saline solution of NKCP was added to human blood immediately after collection to measure laboratory parameters related to coagulation/fibrinolysis. As a result, NKCP treatment decreased both concentrations of thrombin-antithrombin III compound (TAT) and fibrin monomer (FM), showing that it has an anticoagulant action like heparin. Moreover, the concentration of D-dimmer (D-d) was high and that of fibrinogen (Fbn) was low, showing that it has a fibrinolytic effect like alteplase. Different from alteplase, however, the fibrinolytic effect was accompanied by no increase in the concentration of α2 plasmin inhibitor-plasmin compound (PIC), suggesting the fibrinolytic effect is independent of plasmin. As the concentration of NKCP was higher, the effect became larger.

These results demonstrated that the new extract with a molecular weight of 45 KDa from natto culture fluid has an anticoagulant effect and a plasmin-independent fibrinolytic effect on human blood.

iv) Effect of the dried filtrate of natto bacilli culture "NKCP" on blood fluidity
Journal of the Japanese Society of Hemorhelogy Vol. 5 (1), 2002

NKCP was given orally to 13 healthy adults for a week to measure the euglobulin lysis time (ELT) as the indicator for fibrinolysis activity. Doses of 2-4 tablets (equivalent to 250-500 mg of NKCP) daily increased fibrinolysis activity. One healthy adult volunteer, who took 8 tablets (equivalent to 1,000 mg of NKCP) of NKCP daily continuously after meals, showed a remarkably shortened whole blood passage time as determined with a micro channel array flow analyzer (MC-FAN) on Day 7 or later.

Safety

Safety data on natto NKCP

There are currently no reported safety issues or contraindications with NKCP Nattokinase. Being an extract from a traditional food, this product is very safe to take. We do advise, however, that if you have a specific medical problem or if you are taking prescription medications (especially medication relating to thrombosis or blood circulation), you consult your doctor before taking NKCP nattokinase. Over 14 safety studies have been carried out with NKCP, all of which show absolutely no safety problems or toxicity. Eight of these studies are summarized below:

1.	Mutagenicity	Negative
2.	Acute toxicity (rat)	LD50 > 5,000 mg/kg
3.	Subacute toxicity (rat)	Non-toxic-effect dose > 1,325 mg/kg body weight/day male Non-toxic-effect dose > 1,541 mg/kg body weight/day female
4.	Antigenicity (guinea pig)	None (subcutaneous, intravenous)
5.	Excess intake (rat)	Rat models of prolonged coagulation given five times the normal dose through the duodenum showed no bleeding, and NKCP induced no remarkable symptoms in the coagulation system.
6.	Prolongation of bleeding time (rat)	An oral dose of a maximum 300 mg/kg body weight had no effect on the prolongation of the bleeding time.
7.	Long-term intake in humans:	Eight healthy adults were given 750 mg of NKCP daily for 6 weeks and no clinical adverse effects were observed.
8.	Excess intake in humans	23 healthy adults were given 1,250 mg of NKCP daily for 7 days and no clinical adverse effects were observed.

Specification

Spec data on natto NKCP

There are currently no reported safety issues or contraindications with NKCP Nattokinase. Being an extract from a traditional food, this product is very safe to take. We do advise, however, that if you have a specific medical problem or if you are taking prescription medications (especially medication relating to thrombosis or blood circulation), you consult your doctor before taking NKCP nattokinase. Over 14 safety studies have been carried out with NKCP, all of which show absolutely no safety problems or toxicity. Eight of these studies are summarized below:

Appearance	Light yellow powder
Odor	No odor or slight fermented odor
Genetically modified ingredients	none
Moisture	8% or less
Arsenic	1ppm or less
Heavy Metal	10ppm or less
Aerobic Plate Count	3000FU/g or less
Coliform Bacteria	Negative
Protein Produced by B. subtilis natto	90mg/kg or more
Vitmain K2 Content	Negligible
Function Protein Stability	ph 6.0 - 9.0 / <60 td="">
B. subtilis natto cell content	removed / none
Activity	Nattokinase activity was measured by reference to the plasmin substrate (H-D-Val-Leu-Lys-pNA). The activity of 1g is 10 units.
Standard Dose	250-500mg (2-4 tablets) /day
Tablet Ingredients	Proprietary extract of Natto Bacilli Culture, microcrystalline cellulose (bulking agent), resistant dextrin (bulking agent), sucrose esters of fatty acids (emulsifier), silicon dioxide (anticaking agent), shellac (tablet coating). These excipients are completely and are necessary to make the NKCP into tablets.

Activity

Measuring the activity of Natto NKCP

The effectiveness of Natto Extract products is often difficult to compare because there is no standard test for products that help to optimize blood viscosity. Many companies marketing nattokinase compounds measure the enzymatic activity of their product in Fibrinolytic Units (FU), making the assumption that the effectiveness of these enzyme supplements lies solely in the degree with which they can break up fibrin. This is misleading, however, because:

1. Fibrinolysis is just one of the in vitro activities of nattokinase. In vivo, it can have differing activities and functions.
2. Fibrin is an insoluble protein and does not allow formation of uniform suspensions or plates. This makes it unsuitable for a substrate for quantifying enzymatic activity. It should therefore only be used as a substrate for qualitative analysis.
3. Fibrin suspensions or plates have a network structure, and so it is unclear whether the enzymatic activity is associated with the degradation of the network structure or that of fibrin monomers.
4. When fibrin is used as the substrate, the reaction proceeds in a logarithmic manner against the enzyme concentration. Thus, a linear standard curve is not obtained, resulting in an accuracy problem.
5. When fibrin is used as the substrate, reactivity is likely to vary according to the enzymes used. Because there are multiple types of nattokinase, standards for all of these types is necessary. (Current analysis is based on a certain standard product of unknown origin.)

For these reasons, instead of giving the activity of NKCP in FU, Daiwa uses two other methods to determine activity, which results in a much more accurate assessment of their product:

1. Method for determining peptidase activity
   The sample is incubated at 37°C for 5 minutes with the synthetic chromogenic substrate S-2251 (H-D-valyl-L-leucyl-L-lysine-p-nitroanilide dihydrochloride) as substrate and the PNA, which is freed by hydrolysis, is determined by measuring the absorbance at 405 mm. Peptidase activity (unit/gm) is defined 1 unit when 1nmol of pNA per minute is freed from the substrate of 2mM S-2251 at 37°C by 1gm NKCP.
2. Method for determining functional ingredients (antigen measurement method)The natto bacillus-produced protein responsible for NKCP's peptidase activity is purified to prepare antibodies specific for rabbits and mice. The amount of antigen reacting with the specific NKCP antibodies is determined using the sandwich ELISA method. The functional ingredients are represented as μg/g or mg/kg.

Stability

Stability data on natto NKCP

1. Storage Stability

The dried filtrate of B. subtilis natto culture (NKCP Nattokinase) was examined for changes in the content of natto bacillus-produced proteins and peptidase activity during storage. No changes in the content and activity were observed after 6 months of storage at 37ºC or 24 months of storage at room temperature.

Changes in peptidase activity of NKCP powder were ±5% or less when stored at 25ºC or 37ºC for 6 months.

2. Heat Stability

A solution of 1g of NKCP powder in 10ml of 50mM tris–hydrochloric acid buffer (containing 100mM of NaCl, pH 9.0) was used in a heat-stability test for peptidase activity. The NKCP solution, treated under different temperatures and times, was examined for the residual rate of peptidase activity of B. subtilis natto produced protein using a synthetic chromatogenic substrate, S-2251. As shown in the figure below, the peptidase in NKCP was stable when heated up to 60ºC.

Research Papers

Determination of the Thrombolytic Activity of Natto Extract 
Masahiro Msada (Laboratory of Biochemistry, Department
of Bioproduction Science, Faculty of Horticulture, Chiba
University)

Improvement of Blood Fluidity using NKCP, a Dried 
Culture Filtrate of Bacillus Subtilis
Tetsuya Hayashi (Daiwa Pharm), Chie Takahashi
(Microchannel Array Technology Team), and Yuji Kikuchi
(Microchannel Array Techology Team). Journal of 
Hemorheology Research 5 (1), 2002

Research Listing

Foods Pioneer. 1975. “Historical Record of Natto (in Japanese)”, National Cooperative Union of Natto, Tokyo.

Hashima Laboratory of Nihon Bioresearch Inc. 2001.  “Antigenicity Study of NKCP with Guinea Pigs”.

Hayashi T., Takahashi C., and Y. Kikuchi: 2002. “Effect of the dried filtrate of natto bacilli culture "NKCP" on blood fluidity”, Journal of the Japanese  Society of Hemorheology, Vol. 5, pp 43-45.

Hitosugi M., Niwa M. and A.Takatsu: 2000. “Rheologic Changes in Venous Blood During Prolonged Sitting.” Thrombosis Research, pp 409-412, 100.

Hitosugi M., Niwa M. and A.Takatsu: 2001. “Changes in Blood Viscosity by Heparin and Argatroban”, Thrombosis Research, pp 371-374, 104.

Hitosugi M., Tanba M., Koike S., Omura K., Yufu T., Iida N., and S. Tokutome. 2003. “Anticoagulant/ fibrinolytic effect of new natto extract”, Abstracts of  the 26th meeting of the Japanese Society of  Biorheology.

Japan Functional Food Research Association. 1999. “Fibrinolytic Activator of Nattokinase (in Japanese)”, ART Co. Ltd., Tokyo.

Kaketani K., Omura K., and H. Maeda: 2003. “Effect of the natto bacillus culture filtrate "NKCP" on the coagulation/fibrinolysis system”, Submitted to Biorheology.

Kudo T., Uchibori Y., Atsumi K., Numao Y., Miura I., Shitara M. and A. Hashimoto:1978. “Wafarin Antagonism by the fermented soybeans Natto in the Anticoagulant therapy”, Journal of Clinical and  Experimental Medicine, Vol. 104, pp36-38.

Maeda H., Daiwa Pharmaceutical 2003. "Pregnancy and thrombosis, and its prevention",  Medex Journal Vol 46 pp.2-3.

Material Research Center Ltd. 2001. “Evaluation of the thrombolytic activity of NKCP in a rat model of thrombus formation”.

Operation Division, Vivitec Japan, Animal Care Inc. 2000. Study of NKCP on Blood Coagulation Using a Delayed Blood Coagulation Model Duodenal Administration (in situLoop Method)”

Product Safety Labs. 2001. “Acute Oral Toxicity Study in Rats-LD50”

Product Safety Labs. 2001. “Sub-Acute Dietary Toxicity: 90 Day Rodent Study”

Product Safety Labs. 2002. “Thrombolytic Activity and Clotting Potential in Rats”

Sitek Research Laboratories. 2000. “Evaluation of a Test Article in the Salmonella typhimurium/  Escherichia coil Plate Incorporation Mutation Assay in the Presence and Absence of Induced Rat Liver S-9”

Sumi H. 1996. “Invitation to Food Function ~ Functional Food and its Effect (in Japanese)”, pp. 35-45. Sankyo Publication, Tokyo.

Sumi H. 2001. “Nattokinase that is Dramatically Effective against Myocardial Infarction, Cerebral Infarction and Dementia (in Japanese)”, Jitsugyo no Nihon Sha, Ltd. Tokyo.

Sumi H., Hamada H., Nakanishi K. and H.Hiratani: 1990. “Enhancement of the Fibrinolytic Activity in Plasma by Oral Administration of Nattokinase”, Acte  Haematol, pp 139-143, 84.

Sumi H., Hamada H., Tsushima H., Mihara H. and H. Muraki: 1987. “A Novel fibrinolytic enzyme (nattokinase) in the vegetable cheese Natto; a typical and popular soybean food in the Japanese diet”, Experientia, 43, 1110

Tanimura A. 1998. “Handbook on Physiological Activator in Botanical Resources (in Japanese)”., pp. 579-583. Science Forum, Tokyo.

Yamamoto J. 2001. “Anti-fibrinolytic Effect of Nattokinase (in Japanese)”, Research Report 2001  Kobegakuin University.

Yamamoto J. and T. Yamashita. 2002. “The effect of dietary BNPP (Bacillus Natto Productive Protein) administration on experimental thrombolysis model in vivo” Research Report 2001 Kobegakuin University.

Yamashita T. 2001. “In Vivo Evaluation with He-Ne laser-induced thrombolysis model in rat mesenteric microvessel”, Research Report 2001 Kobegakuin  University.

daiwa-pharm.com	Daiwa Pharmaceutical Co. Ltd. in Japan is the developer and manufacturer of NKCP.
dhdeurope.com	DHD Europe is the European branch of Daiwa Pharmaceutical and their site contains good research information.
jafra.gr.jp	Good Japanese site that gives some of the research that has been done with natto nattokinase. ”NKCP” is a dietary supplement which consists mainly of “NKCP”, culture of purified bacillus natto. It’s high in new Nattokinase (Bacillopeptidase), and developed by Daiwa Pharmaceutical Co., Ltd.  NKCP was created from the research of the sticky components Nattokinase which is contained in natto.  We lay in stock the “NKCP” directly from the manufacturer Daiwa Pharmaceutical Co., Ltd.  Per 1 tablet (300mg) Culture of purified bacillus natto(NKCP) – 125mg (Bacillus subtilis var. natto-produced protein - 12.5 micro gram)  NKCP Product Health food containing culture of purified bacillus natto. Ingredients Culture of purified bacillus natto(NKCP), lactose, water-soluble dietary fiber, powdered cellulose, sucrose fatty acid ester, silicon dioxide, shellac (contains soy) Serving 36g (300mg×120 tablets) Nutrition facts ※amount per 1 container (120 tablets) calories 141kcal protein 0.07g fat 1.01g carbohydrate 33.0g sodium 6.8mg Storage Keep away from high temperature, humidity and direct sunlight. Suggested use Take 2 to 4 tablets a day, with water.   Features of NKCP NKCP is a dietary supplement which contains culture of bacillus natto(NKCP). Culture of filtrated bacillus natto was dried with reducing undesirable odor by patented technology of Daiwa Pharmaceutical Co., Ltd. Natto is Japanese traditional food which contains carbohydrate, dietary fiber, a lot of minerals and vitamins. NKCP was created from the research of the sticky components Nattokinase which is contained in natto. NKCP is a collection of useful component which is made by bacillus subtilis var. natto by repeating culture of bacillus subtilis var. natto. It is free from the undesirable odor and viscous texture. Also, the majority of vitamin K2 is removed. The product is maintained to be high in Nattokinase (Bacillopeptidase) constantly. ※The tablets may be irregular in color since NKCP is made from natural ingredient, but it doesn’t affect the quality. ◆Free from the undesirable odor and viscous texture since the majority of vitamin K2, the factor of the smell, is removed. ◆NKCP is maintained to contain specific levels of the protein which has peptide resolution. ◆The production process for this culture of bacillus natto and culture of bacillus natto itself are registered under Patent No. 3532503 in Japan. The culture of bacillus natto(NKCP) is getting high attention from foreign people who are not familiar with natto, and it is expected to be taken in many countries including Asia and Europe. Nattokinase NKCP120 tablets SGD$90.46  (¥7,300)