BOLUOKE - IPlanet Health · 2015-12-19 · known allergy to lumbrokinase, earthworms or corn The above statements have not been evaluated by the FDA. This product is not intended

Is CANADA RNA BIOCHEMICAL INC.

For Circulatory Health

BOLUOKE_ '(Lumbrokinase)

• Hydrolyzes Fibrin • Activates Pieentleett

en

• Degrades Fibrindirr!..

• Inhibits Platelet APP.,

60 CAPSULES

BOLUOKE (Proprietary Earthworm Protein Extract)

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The statements within have not been evaluated by the FDA. The product is not intended to diagnose, treat, cure, or prevent any disease.

CANADA RNA BIOCHEMICAL INC.

"Defining Medicine Of The Future" Local Tel: (604) 273-2233 Fax: (604) 273-2236

Toll Free Tel: (866) 287-4986 www.CANADARNA.COM

Rev. 2014 Oct

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Xa

Va

CC:DAG 1 0 1 Procoagulant Environment

Injury, Inflammation, TT\1F a

TF vii

Extrinsic Pathway

Factor X

Pro Time (PT) (Tenase

Complex) (Cancer Procoagulant a)

CPa

IX, XI, XII

Intrinsic Pathway

Villa APTT

F Prothro mbi n I -i- 2 Fragment 1 +2

Activated Factor X

Prothrombin (Factor II)

(Prothrombinase Complex)

Phospholipid Surface Ca++ co

C Endothelial Till 4,f 4r 4r.foirir Membrane

I bra ne 414 4641.46 41, 446 414

FIBRINOGEN

FM

FIBRIN (13L_Ocic)CLOT)

\_•.. Burst

XI I I

XIlla

I I a Thrombin

(Flopase PhosphatidylSerine Scramblase PhosphatidylCholine Transloc ase)

Hepa ra ns -H AT

Thrc>mbin/Antithrombin

T/AT Complexes

Az Coag Cons 2010 D Berg, MS, Director

© 2001 HEMEX Laboratories

What Is BOLUOKE® ? Scientific name: A proprietary enzyme mixture containing lumbrokinase extracted from

earthworms (Eisenia Fetida)

Main function supports and maintains healthy coagulation activity of the body

Physiological effects already shown in scientific literature activates plasminogen system & hydrolyzes fibrin directly lowers PAI-1, fibrinogen, ESR, and C-RP levels lowers whole blood viscosity & plasma viscosity reduces platelet aggregation

Potential applications Many acute and chronic conditions commonly associated with modern diet and lifestyle often lead to a hypercoagulable blood state, which means the body's fibrinolytic activity is under-functioning relative to the body's pro-coagulation activity. In a hypercoagulable state, these two systems are out of balance and the blood is more prone to forming excessive fibrin. Boluoke® can support the body's fibrinolytic system and bring the whole system back into a more balanced state. Hypercoagulation is not a disease, but merely a blood state that is often associated with many conditions. However, it can contribute to the progression and deterioration of many conditions if it is not brought back into balance. The causes for hypercoagulation may be inflammation, infection, trauma, toxicities, etc., and correcting the underlying causes is the long-term solution.

Major advantages oral preparation; no injection required supported by many scientific and human data does not significantly affect INR, PT or aPTT standardized enzymatic strength excellent safety records and well tolerated

Dosage maximal effect: 2 capsules 3x per day 30 minutes before each meal for 3-6

weeks or as recommended by a physician maintenance: 1 capsule 1-3x per day 30 minutes before each meal

Contra-indications recent surgery, lumbar/arterial puncture, or trauma known aneurysm, GI ulceration/bleeding, or any bleeding disorders concurrent administration of strong anti-platelets like Plavix , Ticlid, etc. known allergy to lumbrokinase, earthworms or corn

The above statements have not been evaluated by the FDA. This product is not intended to diagnose, treat, cure, or prevent any disease

CANADA RNA BIOCHEMICAL INC. WWW.CANADARNA.COM "Defining Medicine Of The Future"

Toll Free: (866) 287-4986 Local: (604) 273-2233 Fax: (604) 273-2236

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Fibrin (Insoluble)

Plasmin Plasminogen

t-PA, Urokinase, Streptokinase

Platelet < Aggregation Inhibition

Fibrinogen

Activation

Boluoke (Lumbrokinase) }

g+ Degradation

+ >

Hydrolyzation

Fibrin Degradation Product (Soluble),

D-Dimer (Insoluble)

Boluoke Mechanism Diagram



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"Boluoke®, The Best Oral Fibrinolvtic Ajient"

A Brief Background:

Boluoke® is a proprietary protein extract with a special ability to balance the body's coagulation system and support a healthy circulation. The main ingredient inside Boluoke® is a group of earthworm enzymes collectively called Lumbrokinase. For many centuries the earthworm has been used in the Far East traditional medical practice and was recorded in the "Ben Cao Gang Mu", the traditional eastern world's pharmacopoeias, as a potent medicine for "Liver Wind uprising and Channel blockage" conditions. In the 1980s Japanese scholar Dr. Hisashi Mihara succeeded in extracting fibrin-dissolving enzymes from earthworms, and named this group of enzymes Lumbrokinase. Since then Korean and Chinese researchers have compiled extensive scientific and clinical data on lumbrokinase. Since early 1990s, modern extracts of earthworms containing lumbrokinase have been available in many countries as a traditional medicine or as a supplement depending on the local health regulations.

Fundamental Studies On Fibrinolysis:

(1) In vitro test shows that 40mg of clot (in 37°C at 9 rpm) can be completely dissolved by 1250u/ml Boluoke® in 30 min. In contrast, Urokinase in the same concentration can only dissolve 18.9% in 30 min., and dissolve 57% in 60 min.; there is still 11% cannot be dissolved at 120min. It shows that the Boluoke® can dissolve clot in vitro with far more efficacy and intensity than Urokinase (see below):

Time Elapsed

(37°C, 9 rpm)

Boluoke8 on 40mg clot Urokinase on 40mg clot

1250 u/ml

125 u/ml 62.5 u/ml 1250 u/ml

300 u/ml 60 u/ml

30 minutes 100% dissolved

66.7% dissolved

27.5% dissolved

18.9% dissolved

N/A N/A

60 minutes N/A N/A N/A 57% dissolved

N/A N/A

120 minutes N/A 100% dissolved

91.3% dissolved

89% dissolved

60% dissolved

31% dissolved



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(2) Researchers induced thrombus in the inferior vena cava in rodents, then gave Boluoke® either 1000u/kg intravenously or 6000 u/kg rectally, and reassess the thrombosis in 2 or 4 hours. Boluoke® was shown to have significant thrombolytic effect both intravenously and rectally compared to the control (see below):

Route of administration

Boluoke® Control Ratio of Animals without

thrombus

Ave. wt. of thrombi

remained

Ratio of Animals without

thrombus

Ave. wt. of thrombi

remained

2 hrs after 10,000 u/kg of Boluoke® intravenously

6/10 (60%) 2.6 +/- 1.29 mg

2/11 (18.18%) 24.36 +1- 6.72mg

4 hrs after 6,000 u/kg of Boluoke® rectally

9/11 (81.81%) 0.87 +/- 0.70mg

1/9 (11.11%) 12.02 +/- 4.85mg

Phamacokinetics & Pharmacodynamics:

(1) Researchers injected emboli made from radioactive 1251 tagged fibrinogen into rabbit jugular veins, and induced pulmonary embolism. The rabbits were then given 2,000 u/kg or 600 u/kg of Boluoke® intra-duodenally or placebo. Blood were taken at 0.5, 1, 2, 3, and 5 hours, and radioactivity were assessed. The Boluoke® group showed significant increase of radioactivity after 3 hours and further increase after 5 hours compared to the placebo group, indicating marked fibrinolysis. The amount of radioactivity also seemed to be dose-dependent (see below):

Radioactivity assessed at:

2,000 u/kg Boluoke® 600 u/kg Boluoke® Control

3 hours 72.9 +/- 12.0 66.7 +/- 13.1 58.1 +/- 8.6 5 hours 70.4 +/- 11.0 72.6 +/- 18.4 52.2 +/- 11.2

(2) Rabbits were given 5,000 u/kg of Boluoke® intravenously, 5 minutes after the euglobulin lysis time (ELT) was shortened from 102 +/- 15 to 15 +/- 2 minutes. When rabbits were given 2,500 u/kg intravenously, 15 minutes after the ELT was shortened from 102 +/- 15 to 42 +/- 17 minutes.

(3) When rabbits were given 4,000 u/kg of Boluoke® orally, ELT started to decrease 3 hours afterwards. At 4 hours post-administration ELT had shorted from 335 +/- 38 to 240 +/- 36 minutes. The ELT dropped to the lowest 8 hours post-administration at 203 +/- 35 minutes. At 12 hours the ELT was back to pre-medication level.

The conclusion of the studies above indicate that Boluoke®'s effect peaks between 5-8 hours after oral administration, thus the optimal way to dose Boluoke® is 3 times daily.



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Animal Toxicity Studies:

1. To determine the LD50 of Boluoke®, researchers chose 40 mice (1/2 males and 1/2 females) weighing 18-20g each, divided them randomly into 4 groups. Each group was given intravenously 200,000 u/kg, 160,000 u/kg, 128,000 u/kg, and 102,400 u/kg respectively, then they were observed for 1 week. The LD50 was determined to be 144,600 u/kg.

2. Water solution of Boluoke® was injected subcutaneously into rodents at 1/4 LD50, 1/10 LD50, and 1/50 LD50, and intravenously into dogs at 1/8 LD50 and 1/50 LD50. This was done once daily for two weeks, and there was no optic or acoustic deficit detected.

3. Rabbits were given intravenous injections of Boluoke® at 500 u/kg, 1,500 u/kg, and 3,800 u/kg. EKG readings were taken prior to injection, immediately after, 10 minutes after, and 90 minutes after injections. There was no detectable EKG change in all groups, except transient tachycardia and hypotension in the rabbits given 3,800 u/kg.

4. 500 u/kg or 1,500 u/kg of Boluoke® were given to rabbits intravenously, and 2,900 u/kg or 18,000 u/kg were given to dogs intravenously. Only the dogs given the 18,000 u/kg dosage had signs of faster and deeper breathing 5 minutes after; all other animals had no respiratory change.

5. Intravenous dose of 2,500 u/kg and 5,000 u/kg were administered to rabbits, and the clotting time was assessed prior, 5 minutes, 15 minutes, 30 minutes, 60 minutes, 90 minutes, and 180 minutes after by the test tube method. There was no significant change detected. Dogs given up to 18,000 u/kg intravenously once daily for 2 weeks also showed no change in clotting time.

6. 80 rodents weighing 160-250g (1/2 males and 1/2 females) were randomly divided into 4 groups. Each group was given subcutaneous Boluoke® at 1/4 LD50, 1/10 LD50, 1/50 LD50, or saline respectively once daily for 2 weeks. There was no significant difference between the treatment and placebo groups in plasma liver enzymes levels, BUN, hemoglobin, and platelet count prior and after the test period. There was also no animal death.

7. 12 dogs weighing 10-17kg (1/2 males and 1/2 females) were randomly divided into 3 groups. Each group received intravenous Boluoke® at 1/8 LD50, 1/50 LD50, or saline respectively once daily for 2 weeks. In the second week, the group receiving higher dose of Boluoke® would experience varying degrees of bowel/urinary incontinence, nausea, vomiting, listlessness, salivation, and loss of balance within one hour of receiving medication; the symptoms would also disappear within that hour. The smaller dosage group and the placebo group did not show the above symptoms. There was no significant difference between the treatment and placebo groups in plasma liver enzymes levels, BUN, hemoglobin, and platelet count prior and after the test period.

8. Mutagenicity and teratogenicity tests showed that Boluoke® did not induce DNA damage/mutation or mitotic aberration. Mice receiving 40 — 1000 u/kg of Boluoke® showed no significant differences in pregnancy weight, fetal growth, abortion rate, still birth rate, and fetal resorption rate compared to the placebo group. There was also no birth defect observed.



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Side Effects:

The earthworm has been used in Traditional Chinese Medicine for a few thousand years, and is considered to be one of the safest medicines in the traditional pharmacopoeias. The earthworm is recorded to have the ability to "eliminate Heat, subdue endogenous Wind, dredge the Channels", and is traditionally used in treating conditions like epilepsy, stroke, hypertension, and pain syndromes. Modern scientific research into earthworm protein extracts and lumbrokinase has also shown them to be extremely safe products. In one of the clinical study on Boluoke® involving 16 hospitals and 1560 patients in China, the overall adverse reaction rate was 1.92% (30 cases). Of the 30 cases, 9 (0.58%) had skin itching, 3 (0.19%) had skin rash, and 18 (1.15%) had nausea or diarrhea; no hemorrhage or major side effect was reported.

Summary:

Boluoke® (lumbrokinase) is the only fully researched oral enzyme preparation on the market which supports the circulatory health, and it shows great promise in maintaining the coagulation/fibrinolysis balance in the body. Coagulation system is a complicated and highly regulated system, and modern lifestyles often tip the balance towards a hypercoagulable blood state. Boluoke® is especially suited for bringing the hypercoagulable blood state into balance.

The use of earthworms to achieve circulatory health in traditional medicine has come a long way. First there was dried earthworms used in traditional oriental medicine decoctions, and then there were ground-up earthworm powders. Later came the granular crude extract of earthworms, and now there is Boluoke®, a nutraceutical product containing purified enzymes extracted from earthworms. Just like omega-3 molecules in fish oil, polyphenols in green tea, and curcumin and turmeric, this group of enzymes called lumbrokinase shall in time be known as the most valuable proteins in earthworms.



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Strength In Plasmin Unit (per gm wet wt.)

Comparison Between Boluoke® (lumbrokinase) and Nattokinase

Fibrinolytic Strength Boluoke® (lumbrokinase) Nattokinase 1,2

Plasmin Unit (CU/g wet wt.) 100 40

t-PA Unit (u/mg) 120,000 12,000** **The nattokinase used in measuring the t-PA activity is from research cDNA source, a

much purer enzyme than the commercially available source from natto extract.

Strength In t-PA Unit (per mg)

140,000 120,000

120,000

100,000

80,000

60,000

40,000

20,000 12,000

.... 0

Boluoke® (lumbrokinase) Nattokinase 1,2

1. Sumi H, Hamada H, Tsushima H, et al. Experientia 1987; 43(10): 1110-1 2. Liu BY, Song HY. Sheng Wu Hua Xue Yu Sheng Wu Wu Li Xue Bao (Shanghai) 2002; 34 (3): 338-40

The enzyme strength varies between products. Please contact individual company for their specifications



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Finally, The Answer You've Been Asking For!

Nattokinase in Whole Blood Viscosity Reduction

**Shear Rate (1/s)

Boluoke® (lumbrokinase) *Subject took 2 caps at baseline

Nattokinase (20,000 FU/gm) *Subject took 4 caps at baseline

Baseline viscosity

(cP)

Viscosity 4 hrs later

Percentage Reduction

(%)

Baseline viscosity

(cP)



(%) 1 15.83 9.67 38.91 16.81 14.01 16.66 2 11.73 8.28 29.41 12.54 10.96 12.60 5 8.61 7.14 17.07 9.28 8.57 7.65

Boluoke® Vs. Nattokinase in Whole Blood Viscosity Reduction

**Shear Rate (1/s)



Baseline viscosity

(cP)



(%)

Baseline viscosity

(cP)



(%) 1 15.28 13.67 10.54 17.01 14.35 15.64 2 11.70 10.74 8.21 12.51 11.34 9.35 5 8.93 8.43 5.60 9.09 8.97 1.32

*The same subject underwent each test with one week of washout period in between. Blood viscosities were check at about 10:30am and 2:30pm. The capsules were taken on an empty stomach after each baseline blood draw. Each capsule of nattokinase contains 36mg of enzyme with a total of 720 FU. **Blood viscosities were performed on Rheolog0 machine of Rheologics, Inc. The particular

Rheolog0 unit used in the experiment appeared to under-estimate viscosities across the board, but was able to consistently reflect changes in viscosity.

Note: From research data available to date on raw materials, we know that Boluoke® (lumbrokinase) is at least 2.5 to 10 times stronger in fibrinolytic strength than commercially available nattokinase. We wanted to see if the finished products would reflect the same potency difference. The above data shows that 2 capsules of Boluoke® containing a total of 40mg lumbrokinase is much more potent than 20 capsules of nattokinase (20,000 FU/gm) containing a total of 720mg nattokinase. That is a potency difference of more than 18 folds!! We understand that this is a simple N=1 trial, and we encourage you to repeat the experiment and find out for yourself. We are sure that you'll find the fibrinolytic strength of Boluoke® is unmatched by any other enzymes on the market.



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Enzymatic Strength Of Boluoke® Vs. Other Enzymes

Product name t-PA Activity / Cap t-PA Activity / mg Relative Strength to Boluoke®

Boluoke® (lumbrokinase) —20mg/cap, 300,000u/cap

Lot#041210

2,597,000u 129,800u 1

AH Nattokinase 100mg/cap, 20,000FU/g

Lot#070403

360,000u 3,600u 1/36

OH Serrapeptidase 20mg/cap, 40,000iu/cap

Lot#B39275006

8,832u 441u 1/294

DH PLA Bacillopeptidase F 250mg/cap Lot#402935

2,906 1 1 u 1/12,900

Notes: All Samples were blinded and tested by Hemostasis ReferenceLaboratory Inc., Hamilton, ON, Canada (Feb. 2011)

Comparison Of Boluoke® With Other Lumbrokinase

Parameter Boluoke® Other Brands

Phase I to III Trials Done Yes No

Pharmacodynamics Data Available Yes No

Pharmacokinetics Data Available Yes No

Toxicity Studies Done Yes No

Teratogenicity Studies Done Yes No

Significantly Affects INR or aPTT No Yes

Also Standardized to t-PA Activities Yes No

Over 20 Years of Safe Clinical Usage Yes No

Contains Corn Ingredients No Yes

Regular Research Update on Lumbrokinase Yes No

Professional Clinical Support Yes Maybe



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Boluoke® (lumbrokinase) FAQ's

1. How long before surgery should patient stop taking Boluoke®? The conservative approach is to stop Boluoke® 1 week prior to surgery. The patient may resume taking Boluoke® 15 days after surgery if no complication or earlier if the physician deems appropriate. Some physicians do recommend patients to stay on a small dose of Boluoke® post surgery in certain high-risk patients.

2. Can Boluoke® be safely taken if the patient is on blood thinning agents? Lumbrokinase has been safely used together with blood thinning agents in various clinical studies, and does not have any adverse interaction nor increase the chance of bleeding other than what is inherent to the blood thinners (Park S, Kye KC, Sumi H, et al. 1989). Boluoke® has been shown not to significantly affect PT and aPTT (thus it does not affect INR). However, patients should always consult their physicians before taking Boluoke® while on any prescriptions.

3. Can the content of Boluoke® capsules be emptied out and taken without the capsules? The capsules used in Boluoke® are enteric coated capsules. They are designed to resist stomach acid and dissolve in the small intestines, because lumbrokinase may be inactivated by stomach acid. Preliminary coagulation tests using Sonoclot® machine (manufactured by Sienco, Inc.) indicate that Boluoke® is still effective when taken without the capsules on an empty stomach. However, it is highly recommended that the patients take Boluoke° in its original capsulated format whenever possible.

4. Can Boluoke® be safely taken with anti-platelet agents? There has been no study specifically looking at this; there has also been no report of adverse interactions to date. However, because Boluoke® can affect platelet aggregation (Zhang GP, Qian RZ, et al. 1998), it should not be used with strong anti-platelets like Plavix or Ticlid unless under the supervision of a physician.

5. Can Boluoke® be safely taken with aspirin? Boluoke° has been shown to be safe when taken with aspirin in clinical studies (Wang XL, Yan DC. 2000; Wang H, Yan DC, et al. 2000). However, patients should consult with their physicians before taking Boluoke® with aspirin or other NSAIDs.

6. Can Boluoke® be used on pregnant women? Teratogenicity studies of Boluoke° on animals showed no effect on pregnancy weight, fetal growth, abortion rate, still birth rate, and fetal resorption rate in mice compared to the placebo group. There was also no birth defect detected. However, pregnant patients should not take Boluoke° unless it is done under a physician's approval and supervision.



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Boluoke® (lumbrokinase) FAQ's (cont'd)

7. What are the contra-indications for taking Boluoke®? Contra-indications for Boluoke® are: allergy to lumbrokinase or earthworm; recent surgery; pre-surgery; lumbar puncture or arterial puncture; trauma; high risk aneurysm; active internal bleeding or GI ulceration; any other bleeding disorders/tendencies. People who are taking any anti-coagulants (e.g. heparin, coumadin) or anti-platelet drugs (e.g. Plavix or Ticlid) should consult their physicians before taking Boluoke®.

8. Does Boluoke® have any side effects? The earthworm has been used in Traditional Chinese Medicine for a few thousand years, and is considered to be one of the safest medicines in the traditional pharmacopoeias. In one of the largest clinic trial of Boluoke® involving 16 hospitals and 1560 patients in China, the overall adverse reaction rate was 1.92% (30 cases). 0.58% had skin itching, 0.19% had skin rash, and 1.15% had nausea or diarrhea; no hemorrhage or major side effect was reported.

9. Does Boluoke® affect ESR or C-RP? What about PAI-1(Plasminogern Activator Inhibitor 1)? Boluoke® has been shown to reduce ESR up to 30% in 4 weeks and C-RP up to 40% in 3 weeks. It has also been shown to reduce PAI-1 in a human clinical trial. PAI-1 is believed to be a risk factor for the development of Type 2 Diabetes.

10. How fast and how much can Boluoke® lower the patient's plasma fibrinogen level? On average, a reduction rate of 10-20% (or more) in 4 weeks can be expected when patients are on the full dosage of 2 capsules three times daily. Boluoke® does not reduce plasma fibrinogen level to below normal. Fibrinogen is an inflammatory marker, and physicians should seek the underlying causes of the inflammation and correct them, and not simply treating the elevated fibrinogen level.

11. How fast and how much can Boluoke® lower the patient's whole blood viscosity? Preliminary data shows that Boluoke® could reduce low-shear whole blood viscosity up to 38% in 4 hours (using Rheolog® machine by Rheologics). However, in some patients with high baseline whole blood viscosity, the viscosity at 4-6 hours after oral dosing may increase temporarily (possibly due to increased generation of FDP & D-Dimers). If the patient continues with the dosage (1-2 caps tid), the viscosity should come down nicely at 24 hours.



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12. Can Boluoke® be taken with other enzyme products? There is a theoretical possibility that Boluoke® may be cleaved and rendered ineffective by other proteolytic enzymes, so we currently do not recommend dosing Boluoke with another proteolytic enzyme product. If you do have to take other such enzyme products while on Boluoke®, try to space more time between the dosing of the two enzyme products (i.e. at least 2 hours).

13. What is lumbrokinase's effect on FDP (Fibrin Degradation Product) or D-Dimer? FDP and D-Dimer levels have been shown to increase significantly from the baseline within 24-48 hours of lumbrokinase administration in healthy and hyperfibrinosis individuals. FDP and D-Dimer are indications of fibrinogen and fibrin breakdown in the body.

14.1 have heard that Boluoke® can help Peyronie's disease, is it true? There is no evidence indicating that Boluoke® or other similar enzymes have any effect in helping Peyronie's disease. Knowing the mechanisms of Boluoke®, it is very unlikely that it will be helpful for Peyronie's disease, thus it is not recommended for those patients.

15. What about uterine fibroids, is Boluoke® useful? Again, this is another myth that's been propagated on the Internet. Boluoke® is not likely to help those patients unless there is coexisting hypercoagulation.

16. Can Boluoke® be used to help lower cholesterol? There is some evidence indicating that lumbrokinase may have a mild cholesterol lowering effect. However, we believe there are many other more effective ways to lower the cholesterol. Thus, Boluoke® should not be recommended for cholesterol lowering purpose but for the overall health of the circulatory system.

17. Can Boluoke® be used to replace coumadin? Boluoke® is a nutraceutical product and should not be used to replace coumadin. However, if a patient refuses to take coumadin as recommended, then the physician shall find the best alternative for the patient. Regardless what alternatives the physician decides on, the patient should be properly monitored. If Boluoke® was chosen as the best alternative, the patient should be checked for anti-plasmin, prothrombin fragment 1+2, thrombin/antithrombin complex, and soluble fibrin monomer on a regular basis in order to make sure the thrombosis risk is minimal. A discussion with specialized hematology lab as to what might be the best way to monitor the patient's status is highly recommended.



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18. What is the suggested protocol for taking Boluoke®? Boluoke® is best taken under a physician's guidance in accordance to the patient's individual condition. For most chronic conditions and maintenance, 1 capsule once to three times daily on empty stomach should be sufficient. In acute conditions Boluoke® can be taken at the dose of 2 capsules three times daily on an empty stomach for 3 to 6 weeks or as long as the doctor recommends.

19. What kind of tests can be used to see if patients are on appropriate doses of Boluoke®? Tests like anti-plasmin, prothrombin fragment 1+2, thrombin/antithrombin complex, or soluble fibrin monomer levels are some possible tests to help physician determine if patients are on appropriate doses of Boluoke®. We have also found that the gbACT+ test performed on Sonoclot® machine (manufactured by Sienco, Inc.) is a very helpful tool for in-office monitoring of patient status. However, a discussion with specialized hematology lab as to what might be the best way to monitor patient status is highly recommended.

20. How quickly does Boluoke® start to act on the body and how to best maintain the maximal effect? Pharmacokinetic studies showed that the fibrinolytic effect of Boluoke® kicks in about 3 hours and peaks at 6-8 hours after oral administration. At about 12 hours after oral dosing, the fibrinolytic activity is back to baseline level. Thus, the best way to obtain and maintain maximal benefit from Boluoke® is to take each dose about 7 to 8 hours apart.

21. How is Boluoke® different from other products that contain lumbrokinase? Boluoke® is the only lumbrokinase that has been through all phases of clinical trials and its enzymatic strength is standardized against tPA and urokinase activities. Other lumbrokinase products currently on the market often are citing Boluoke®s credentials and research, and are not disclosing the enzymatic strength of their lumbrokinase. On the raw material market, the price of lumbrokinase can vary by up to 15 folds, and the enzymatic strengths of various lumbrokinase also differ greatly. In addition, lumbrokinase is a preparation containing multiple enzyme fractions, and the extraction and purification method determines the composition of the various enzyme fractions. Thus different extraction method will produce different sub-fractions of lumbrokinase. This is the reason why Boluoke® does not significantly change prothrombin time (PT) or activated partial thromboplastin time (aPTT), while other lumbrokinase sources may significantly alter PT or aPTT as shown in some studies (Jin L, Jin H, Zhang G, Xu G. 2000).



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REFERENCES

Fan Q, Wu C, et al. Some features of intestinal absorption of intact fibrinolytic enzyme III-1 from Lumbricus rubellus. Biochem Biophys Acta, 2001; 1526(3): 286-92

Gao Y, Qin MZ. Lumbrokinase in treatment of patients with hyperfibrinogenemia of coronary atherogenesis disease. Journal of Capital University of Medical Sciences, 1999; 4(20)

Gong B, Wu XY. Observation of using Baiao lumbrokinase capsules to treat ischemic cerebrovascular accident with hyperlipidemia. Capital Medicine, 2000; 7(12): 39

Guo ZF, Liu XX. Observation of treating ischemic cerebrovascular accident with Baiao lumbrokinase capsules. Capital Medicine, 2000; 7(11): 45

Huang ZD, Li ZW, Zhang WX. Lumbrokinase in treating cerebral infarction. Chinese Journal of New Drugs and Clinical Remedies, 2000; 6(19): 453-455

Jie WH. Clinical observation of treating unstable angina in seniors with lumbrokinase. Capital Medicine, 2000; 7(10): 37

Jin L, Jin H, Zhang G, Xu G. Changes in coagulation and tissue plasminogen activator after the treatment of cerebral infarction with lumbrokinase. Chinese Journal of Microcirculation, 2000; 23: 213-218.

Jing LR, Xu GZ. Dynamics of fibrinolysis and hemostasis in ischemic stroke patients, and the effects of lumbrokinase on those dynamics. Chinses Journal of New Drugs and Clinical Remedies, 1999; 18(1): 48-49

Liao RH. Analytical report of treating 30 patients of ischemic cerebrovascular disease with Panford lumbrokinase and nimodipine. Strait Pharmaceutical Journal, 1997; 9(3): 25-26

Liu J, Zhou LM, Ren Y. Lumbrokinase capsule vs Salvia miltiorrhiza tablet in treating coronary disease with angina pectoris. Chinese Journal of New Drugs and Clinical Remedies, 1999; 1(18): 17-19

Mihara H, Sumi H, Mizumotoh, et el. Oral administration of earthworm powder as a possible thrombolytic therapy ed In: Tamkak, Recent advance in thrombosis and fibrinolysis. Hapan Academic Press, 1996, 287.

Mihara H, Sumi H, Yoneta T, Mizumoto H, et al. A novel fibrinolytic enzyme extracted from the earthworm, Lumbricus rubellus. Jpn J Physiol, 1991; 41: 461-72.

Mihara H, Yoneta T, Sumi H, et al. A possibility of Earthworm powder as therapeutic agent for thrombosis. Thromb Haemost, 1989; 62: 545.



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REFERENCES (coned)

Mihara H, Sumi, H, Akazawa K, et al. Fibrinolytic enzyme extracted from the earthworm. Thromb Haemost, 1983; 50(1): 258.

Pan SY. Treating Blood-Stagnation type angina with Baiao lumbrokinase. Capital Medicine, 1998; 9(5):40-41

Pang SQ, Ding MC, Xie SP, et al. A clinical study of therapeutic effectiveness in treating ischemic cerebrovascular disease with Lumbrokinase (Boluoke). Chinese Journal of Neurology and Psychiatry, 1993; 26(4): 229.

Park S, Kye KC, Sumi H, et al. Fibrinolytic activity of the earthworm extract. Thromb Haemost, 1989; 62: 545.

Park Y, Ryu E, Kim H, Jeong J, et al. Characterization of antithrombotic activity of lumbrokinase-immobilized polyurethane valves in the total artificial heart. Artif Organs,1999; 23: 210-4.

Qi W, Yu XB, et al. Effects of lumbrokinase on blood rheology of seniors with coronary arterial diseases. Chinese Journal of Microcirculation, 2000; 1(10): 55

Ryu GH, Park S, Kim M, et al. Antighrombogenicity of lumbrokinase-immobilized polyurethane. J Biomed Mater Res, 1994; 28: 1069-77.

Tang Y, Liang D, et al. Crystal structure of earthworm fibrinolytic enzyme component a: revealing the structural determinants of its dual fibrinolytic activity. J Mol Biol, 2002; 32(1): 57-68

Wang H, Yan DC, et al. Effects of Baiao lumbrokinase on plasma endothelin concentration and left ventricular function. Capital Medicine 2000; 7(1): 44.

Wang LH, Yao W. Vermis kinase (Boluoke) therapeutic evaluation to cerebral infarction and influence to blood rheology. Journal of Practical Medical Technology, 1998; 5(2).

Wang R, Chen Q, Fang XQ, et al. Effects of Fleroxacin combined with urokinase or earthworm kinase on bacterial biofilm. Acta Pharmaceutica Sinica, 1999; 9(34):662-665

Wang XL, Yan DC. Clinical observation of using Baiao lumbrokinase capsules to treat patients with coronary artery disease secondary to diabetes mellitus. Capital Medicine, 2000; 7(8): 38



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REFERENCES (coned)

Wu XQ, Wu C, et al. Immobilized earthworm fibrinolytic enzyme III-1 with carbonyldiimidazole activated agarose. Protein and Peptide Letters, 2002: 9(1): 75-80

Yang GR. Treating unstable angina with Baiao lumbrokinase capsules. Capital Medicine. 1999; 3(6): 30

Zhang GP, Jin HM, Zhang M, et al. Anticoagulative and fibrinolytic effects of lumbrokinase and their relation to tissue plasminogen activator. Chinese Journal of Geriatrics, 1998; 6(17): 366-368

Zhang GP, Qian RZ, et al. Experimental observation of the inhibitory effects of lumbrokinase on platelets. National Medical Journal of China, 1998; 5(78): 394-95

Zhang HY. Clinical evaluation of treating acute ischemic cerebrovascular disease with lumbrokinase. Capital Medicine, 2000; 7(3): 45-46

Zhang JQ, Jing CH. Clinical evaluation of using Baiao lumbrokinase in the treatment of central retinal vein occlusion. Capital Medicine, 1999; 6(2): 51

Zhang LP, et al. Preventative effects of lumbrokinase on experimentally induced emboli. Chinese Journal of Hemorheology, 1995, 11(10): 679-680

Zhang NH. Blood rheological changes of coronary artery disease patients prior and after lumbrokinase treatment. Chinese Journal of Hemorheology, 1999; 1(9): 63

Zheng HJ, Xu JM, Huang ZH. Lumbrkinase capsule vs ticlopidine in treating coronary artery disease with angina pectoris. Chinses Journal of New Drugs and Clinical Remedies, 2000; 5(19): 406-408

Zheng Z, Lin JN, et al. Observation of 20 unstable angina patients treated with Baiao lumbrokinase capsules. Capital Medicine, 2000; 7(6): 38

Zhao J, Fan R, et al. Assay of lumbrokinase with a chromophoric substrate. Protein and Peptide Letters, 1997; 4(6): 409-14

Zhou M, Wang FQ. The effects of lumbrokinase on blood rheology in patients with sudden hearing loss. Journal of Chinese Microcirculation, 2000; 3(4): 177-178

Zhou XD, Zhao RX, Zheng XR, Zhang H. Observation of Boluoke (lumbrokinase) treatment in the recovery of motor function after cerebral infarction. Chinese Journal of Western and Chinese Medicine Cooperation, 1997; 12 (17).



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COAG 101 FIBRINOLYSIS & INHIBITORS

Xa

Protein

Protein C APC

Ex Pat ► way

2) HAS

TFPI Xi

1) B2 -GP -I or

Pig

S Su Hep Ann exi n II

to

I :404644"ir 4•17 vhcisphatidyl Seri nc.. (negative charg)

Endothelial Cell Membrane phosphaticlylChnline (rmutral charg0

TF VII

Thrombin

T'M ThromboM o du li n

Factor X

mla•MMI ON.

Thrombin (11a)

(Heparin/AT) HAT

IX, XI, XII

Intrinsic Pathway

APC I PS

H a *TAT

JAT Aritiffirorribin

Lp(a) 5) Plasminogen

+tPA PAI-1

► VI I li

Thrombin/Antithrontin

P7IN

Az Coag 2010

Complexes

VIII a

Boluoke4, Simply The Best!!

Microcirculation is often compromised in patients with chronic conditions. A healthier circulation goes a long way in making any treatment plan more effective and in helping the patients get better sooner.

When you recommend a second grade enzyme product for you patients, you are putting their health in potential jeopardy and your professional reputation on the line.

Your Patients' Health Your Reputation

Trust Nothing Less



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Phase III Clinical Trial Summary of Boluoke Capsule in Treating Ischemic Cerebrovascular Disease

Cerebral infarct is one of the most commonly seen cerebrovascular diseases; it has an abrupt onset, a high morbidity and affects people's psychosomatic health mostly. Up to date, no oral medicine with the effect of tissue-typed plasminogen activator (t-PA) has been found, though there are many drugs for treatment of cerebral infarct. Boluoke capsule is a multi-component pure-natural medicine extracted from earthworms, and includes at least two kinds of enzymes: plasminogen activator and plasmin.

Since 1990, affiliated Xuanwu Hospital of Capital Medical College, Jiangxi Provincial People's Hospital and No.2 Affiliated Hospital of Jiangxi Medical College have studied the clinical manifestations and the blood rheology changes in 453 patients with ischemic cerebrovascular disease by randomized double-blinded method.The results indicated that the total effective rate is 93.73% and the significant response rate is 73.60%. Organized by Chinese Medical Society, a collaborative group of 16 hospitals has performed a phase III clinical trial of Boluoke capsule in treating ischemic cerebrovascular disease from June 1992 to December 1993. Totally, 1560 patients were studied according to "the censorship and approval regulations of new drugs "to further verify the clinical efficacy and investigate the adverse effects. The total effective rate is 88.21% and the significant response rate is 68.91 %. Several hospitals also studied the changes in blood rheology. These results all showed that Boluoke capsule is a promising new anti-thrombotic agent with a definite efficacy and no obvious toxic/adverse effects, worthy of extensive applications.

Patients and Methods

1) Case selection: All cases in our trial were diagnosed according to the Diagnostic Criteria of the Second National Symposium on Cerebrovascular Diseases (China). They all had various degrees of hemiplegia and infarct foci shown on CT scan. The clinical efficacy of 1560 patients was investigated according to the unified therapy protocol. There were 1001 males and 559 females; the mean age was 52.38 +/- 8.13. The majority of patients took Boluoke capsule within one month from the onset (see Table 1).

2) Administration method: Two Boluoke capsules were taken orally half an hour before meal each time, three times a day for 21 days. During this period, other drugs that can dilate cerebral vessels, affect blood rheology and deoppilate the blood vessels were prohibited.

3) Appraisal of the clinical efficacy: The clinical neural deficit before and after the administration was scored by "The score criteria of clinical nervous function deficiency degree in patients with stroke" passed by the Second National Symposium on Cerebrovascular Diseases. Score 0-15 was defined mild; score 16-30 moderate and score 31-45 severe." Beijing score criteria of clinical efficacy in patients with stroke was adopted in the finalappraisal of efficacy (score declined more than 90% was defined basically recovered; score declined from 46% to 89% significantly improved, score declined from 18% to 45% improved; score declined less than 18% or increased less than 18% no changed and deficiency score increased more than 18% deteriorated). Some hospitals also studied the changes in fibrinogen and euglobulin lysis time. The blood was sampled at 8:00-8:30AM and the sampled site was ante-cubital vein.

4) Data processing: t-test and X2

test were applied with TheStatistic Software of The Capital Medical College Mathematics Faculty. All data were processed by IBM-PC.


(cont' d)

Results

The results indicated that the total effective rate was 88.21 %, and the significant response rate was 68.91 % (see Table 2). The results of those hospitals performed fibrinogen and euglobulin lysis time tests showed that fibrinogen and euglobulin lysis time were significantly reduced and the blood rheology was significantly changed after using Boluoke capsule (see Table 3 and Table 4). The adverse effects were very rare (see Table 5).

Discussion

Using earthworms as medicine was recorded in the Chinese traditional medicine masterpiece entitled Ben Cao Gang Mu (The Catalog of Herbal Medicine). It was deemed that the earthworm has the nature of warding off "wind" and unblocking the body's meridians and channels, and can be used to treat hemiplegia or pain syndromes due to "evil" blockages. In the 1983's international symposium of thrombosis and hemostasis, a Japanese scholar reported that the extract of earthworms had the effect of thrombolysis and denominated it "Lumbrokinase". In 1986, Korea Drugs Administration Bureau approved the manufacture of Lumbrokinase. Since then this enzyme has come into the market of many countries abroad and has been applied extensively. The trade name of Lumbrokinase in Hong Kong is "the Hart of Dragon". Boluoke capsule is a multi-component enzyme preparation and has the analogous component of t-PA, which can lyse the thrombi in vivo. There are at least two mechanisms: one is that Boluoke has the effect of plaminogen activator and the other is that it has special affinity to fibrin and thus degrades fibrin rapidly.

Because Boluoke capsule is an enzyme preparation, it is best taken half an hour before meals in order to reduce the effect of gastric acid. The low gastric volume and gastric acid enables the enzyme to be transported to intestinal tract as soon as possible.

In recent years, with further study on bioengineering technology, how large molecules such as protein penetrates the biomembrane has been extensively investigated. Fifteen years ago, Dr. Adiwl in US suggested that protein could be absorbed in vivo as peptides. In 1989, a Japanese professor verified this point by experiment. Many studies have found the general phenomena of trans-membrane transportation of protein and have elucidated theoretically that protein and enzyme can be transported transmembranely to all parts of the body and exert their normal physiological functions. Biophysics Institute of Chinese

Academy of Sciences has made animal models to verify the thrombolysis effect of Boluoke. 125j_ Labelled fibrinogen was intermingled when the thrombi were formed on the rabbit model for acute pulmonary artery embolism. Then Boluoke was given via duodenum and radioactive intensity was measured at the time of half-an hour, 1 hour, 2 hours and 5 hours post-administration. The result demonstrated that the radioactivity significantly increased 3 hours and 5 hours after administration, which means the effect of thrombolysis was significant 4-8 hours after administration. A Japanese professor also reported that the replicated venous thrombi of Beagle dog were dissolved overtly after oral administration of Lumbrokinase.

General Information of the Patients Table 1

Hospital No. of Patients Age Condition of the Illness

Mild Moderate Severe

Dur. of Illness Bef.

Trtmt (# of Days)

Pretreatment

Score Total Male Female 1 Beijing Xuanwu Hospital 210 126 84 61.51 ± 2.54 110 85 15 2.32 ± 2.54 17.16 ± 8.23

2 No. 2 Affiliated Hospital of Jiangxi Medical College 171 112 59 63.3 ± 9.13 89 73 9 3.61 ± 3.17 16.51 ± 7.11

3 Jiangxi Peoples Hospital 130 87 43 64.35 ± 9.83 105 20 5 26.72 ± 78.22 10.42 ± 7.50

4 Zhongshan Affiliated Hospital of Shanghai Medical College 50 27 23 66.49 ± 8.49 24 23 3 31.35 ± 37.59 15.51 ± 7.42

5 Affiliated Hospital of Tianjin Medical College

33 26 7 61.73 ± 9.38 10 17 6 26.72 ± 78.22 19.15 ± 11.36

6 Affiliated Hospital of West China Medical College 60 40 20 60.14 ± 7.78 28 27 5 6.10 ± 6.70 18.20 ± 9.15

7 Nanjing Brain Hospital 43 29 14 63.4 ± 8.78 33 7 3 13.74 ± 21.27 8.84 ± 9.75 8 Sichuan Peoples Hospital 50 39 11 66.2 ± 9.99 26 21 3 9.74 ± 23.76 16.2 ± 8.86

9 Affiliated Hospital of Chongqing Medical College 42 25 17 63.91 ± 9.29 29 12 1 6.21 ± 6.60 10.38 ± 8.77

10 No. 731 Hospital of Space Ministry 183 124 59 58.3 ± 9.52 120 44 19 7.23 ± 28.72 15.73 ± 8.89 11 No. 1 Handan Hospital 128 77 51 59.41 ± 8.67 95 31 2 2.72 ± 3.03 12.16 ± 7.35

12 Affiliated Hospital of Handan Second Medical School 97 53 44 62.00 ± 6.50 48 48 1 1.83 ± 2.17 16.67 ± 7.77

13 Handan Central Hospital 83 55 28 59.10 ± 8.26 46 33 4 3.33 ± 5.94 16.23 ± 8.14

14 Central Hospital of Handan Mineral Bureau 48 30 18 64.15 ± 5.28 28 20 0 1.50 ± 1.01 15.25 ± 7.13

15 An'yang Peoples Hospital 132 87 45 50.26 ± 8.07 80 42 10 2.97 ± 3.77 15.04 ± 9.87 16 Tangshan Workers' Hospital 100 64 36 61.31 ± 8.04 68 26 6 8.92 ± 10.15 13.54 ± 8.97

Total 1560 1001 559 62.38 ± 8.13 939 529 92 9.93 ± 14.20 13.59 ± 8.20

Clinical Response to Boluoke® (1560 cases total) Table 2

Hos itals p # of Patients Basically

Recovered Significantly

Improved Improved No Change Significant Response Rate

Total Effective Rate

1 Beijing Xuanwu Hospital 210 28 (13.33%) 98 (46.67%) 44 (20.95%) 40 (19.05%) 126 (60%) 170 (80.95%)

2 No. 2 Affiliated Hospital of Jiangxi Medical College 171 64 (37.43%) 59 (34.50%) 37 (21.64%) 11 (6.43) 123 (71.93%) 160 (93.57%)

3 Jiangxi People's Hospital 130 17 (13.08%) 51 (39.23%) 42 (32.31%) 20 (15.38%) 68 (52.31%) 110 (84.62%)

4 Zhongshan Affiliated Hospital of Shanghai Medical College 50 2 (4.00%) 16 (32%) 25 (50%) 7 (14%) 18 (36%) 43 (86%)

5 Affiliated Hospital of Tianjin Medical College 33 4 (12.12%) 14 (42.42%) 10 (30.3%) 5 (15.15%) 18 (54.55%) 28 (84.85%)

6 Affiliated Hospital of W. China Medical College 60 8 (13.33%) 25 (41.67%) 15 (25%) 12 (20%) 33 (55%) 48 (80%) 7 Nanjing Brain Hospital 43 21 (48.84%) 12 (27.91%) 4 (9.3%) 6 (13.95%) 33 (76.74%) 37 (86.05%) 8 Sichuan People's Hospital 50 4 (8%) 34 (68%) 9 (18%) 3 (6%) 38 (76%) 47 (94%)

9 Affiliated Hospital of Chongqing Medical College 42 18 (42.86%) 13 (30.95%) 4 (9.52%) 7 (16.67%) 31 (73.81%) 35 (83.33%) io No. 731 Hospital of Space Ministry 183 87 (47.54%) 76 (41.53%) 7 (3.83%) 13 (7.1%) 163 (89.07%) 170 (92.9%)

11 No. 1 Handan Hospital 128 19 (14.84%) 73 (57.03%) 20 (15.63%) 16 (12.5%) 92 (71.88%) 112 (87.5%)

12 Affiliated Hospital of Handan 2'd Medical School 97 22 (22.68%) 44 (45.36%) 23 (23.71%) 8 (8.25%) 67 (69.07%) 89 (91.75%)

13 Handan Central Hospital 83 26 (31.33%) 41 (49.40%) 8 (9.64%) 8 (9.64%) 67 (80.72%) 75 (90.36%) 14 Central Hospital of Handan Mineral Bureau 48 11 (22.92%) 19 (39.58%) 11 (22.92%) 7 (14.58%) 30 (62.5%) 41 (85.42%)

15 An'yang People's Hospital 132 29 (21.97%) 71 (53.79%) 21 (15.91%) 11 (8.33%) 100 (75.76%) 121 (91.67%)

16 Tangshan Workers' Hospital 100 21 (21%) 48 (48%) 21 (21%) 10 (10%) 68 (68%) 90 (90%) 17 Total 1560 381 (24.42%) 694 (44.49%) 301 (19.29%) 184 (11.79%) 1075 (68.91%) 1376 (88.21%)

**No. of patients that have deteriorated or died during the trial: 0% out of the 1560 cases**

Fibrinogen and Euglobulin Lysis Time Changes Table 3

Hospital

[§ P<0.05 , * P<0.01]

No. of

Pts

Fibrinogen Level (mg/di) Euglobulin Lysis Time (Minute)

Before Treatment After Treatment Before Treatment After Treatment

1 Beijing Xuanwu Hospital 210 355.86 +/- 55.98 294.77 +/- 45.15 * 196.89 +/- 70.24 153.40 +/- 48.00*

2 Jiangxi People's Hospital 130 414.39 +/- 159.15 356.50 +/- 103.88* 242.55 +/- 64.56 207.85 +/- 56.54*

3 Zhongshan Affiliated Hospital of Shanghai Medical College 50

352.14 +/- 307.31 208.38 +/- 138.14* N/A N/A

4 College Affiliated Hospital of Chongqing Medical

42 N/A N/A 176. 15 +/- 40.58 129.23 +/- 59.51§

5 No. 731 Hospital of Space Ministry 183 448.47 +/- 124.91 381.56 +/- 123.35* 194.16 +/- 71.80 156.83 +/- 55.36*

6 No. 1 Handan Hospital 128 304.03 +/- 39.55 295.78 +/- 34.35* 138.08 +/- 34.49 119.06 +/- 24.40*

7 Affiliated Hospital of Handan Second Medical School 97

391.54 +/- 70.66 311.71 +/- 61.34* 152.80 +/- 33.73 130.30 +/- 23.81*

8 Handan Central Hospital 83 376.87 +/- 56.32 323.48 +/- 58.91* 151.93 +/- 44.46 131.37 +/- 32.95*

9 Central Hospital of Handan Mineral Bureau 48 358.75 +/- 82.42 314.79 +/- 50.74* 165.81 +/- 33.99 130.46 +/- 24.53*

10 An'yang People's Hospital 132 390.26 +/- 54.95 334.25 +/- 39.67* 224.11 +/- 41.57 214.95 +/- 42.04*

11 Tangshan Workers' Hospital 100 330.05 +/- 83.28 269.88 +/- 58.89* N/A N/A

Blood Viscosity, Plasma Viscosity, Platelet Aggregation before and after Treatment Table 4

Hospital (*P<0.01, § P<0.05)

# of

Pts

Whole Blood Viscosity Plasma Viscosity

Pretrtmt After Trtmt

PAgT Pretrtmt After Trtmt Pretrtmt After Trtmt

1 Beijing Xuanwu Hospital 210 5.60 +/- 0.59 5.18 +/- 0.54* 1.83 +/- 0.12 1.72 +/- 0.09* 67.08 +/- 17.52 46.20 +/- 14.00 2 Zhongshan Affiliated Hospital of Shanghai

Medical College 50 7.28 +/- 1.64 6.48 +/- 1.47* 1.71 +/- 0.15 1.56 +/- 0.22* N/A N/A

3 Nanjing Brain Hospital 43 7.68 +/- 2.39 6.98 +/- 1.85 § 1.90 +/- 0.80 1.85 +/- 0.58 § N/A N/A 4 No. 721 Hospital of Space Ministry 183 9.35 +/- 2.51 8.30 +/- 1.72* 1.77 +/- 0.21 11.67 +/- 0.12* 61.14 +/- 18.12 51.70 +/- 18.89 5 No. 1 Handan Hospital 128 8.95 +/- 1.13 8.15 +/- 0.77* 1.81 +/- 0.11 1.70 +/- 0.08* 67.89 +/- 6.72 63.63 +/- 8.65

6 Central Hospital of Handan Mineral Bureau 48 6.77 +/- 0.87 6.20 +/- 0.72* 1.85 +/- 0.12 1.66 +/- 0.11* 71.25 +/- 11.33 65.06 +/- 4.75

Affiliated Hospital of Handan Secondary Medical School 97 7.02 +/- 1.27 6.02 +/- 1.21* 1.83 +/- 0.12 1.70 +/- 0.10* 70.01 +/- 7.00 64.98 +/- 5.78

8 Handan Central Hospital 83 7.80 +/- 1.65 6.92 +/- 1.45* 1.87 +/- 0.12 1.72 +/- 0.13* 67.11 +/- 9.59 61.28 +/- 5.2 9 An'yang People's Hospital 132 8.27 +/- 1.73 8.19 +/- 1.26* 1.70 +/- 0.11 1.57 +/- 0.15* 57.49 +/- 22.18 47.58 +/- 22.1 # Tangshan Worked Hospital 100 4.42 +/- 0.56 4.03 +/- 0.41* 1.69 +/- 0.14 1.00 +/- 0.09* 73.08 +/- 13.39 60.10 +/- 10.

Symptoms No. of Cases Percentage Skin Itching 9 0.58%

Skin Rash 3 0.19%

Nausea 18 1.15%

Out of 1,560 cases: Total Cases of Adverse Reactions

30 1.92%

Adverse Reaction Rates (out of 1560 cases) Table 5


Cerebral infarct is one of the most commonly seen cerebrovascular diseases; it has an abrupt onset, a high morbidity and affects people's psychosomatic health mostly. Up to date, no oral medicine with the effect of tissue-typed plasminogen activator (t-PA) has been found, though there are many drugs for treatment of cerebral infarct. Boluoke capsule is a multi-component pure-natural medicine extracted from earthworms, and includes at least two kinds of enzymes: plasminogen activator and plasmin.

Since 1990, affiliated Xuanwu Hospital of Capital Medical College, Jiangxi Provincial People's Hospital and No.2 Affiliated Hospital of Jiangxi Medical College have studied the clinical manifestations and the blood rheology changes in 453 patients with ischemic cerebrovascular disease by randomized double-blinded method.The results indicated that the total effective rate is 93.73% and the significant response rate is 73.60%. Organized by Chinese Medical Society, a collaborative group of 16 hospitals has performed a phase III clinical trial of Boluoke capsule in treating ischemic cerebrovascular disease from June 1992 to December 1993. Totally, 1560 patients were studied according to "the censorship and approval regulations of new drugs "to further verify the clinical efficacy and investigate the adverse effects. The total effective rate is 88.21% and the significant response rate is 68.91 %. Several hospitals also studied the changes in blood rheology. These results all showed that Boluoke capsule is a promising new anti-thrombotic agent with a definite efficacy and no obvious toxic/adverse effects, worthy of extensive applications.

Patients and Methods

1) Case selection: All cases in our trial were diagnosed according to the Diagnostic Criteria of the Second National Symposium on Cerebrovascular Diseases (China). They all had various degrees of hemiplegia and infarct foci shown on CT scan. The clinical efficacy of 1560 patients was investigated according to the unified therapy protocol. There were 1001 males and 559 females; the mean age was 52.38 +/- 8.13. The majority of patients took Boluoke capsule within one month from the onset (see Table 1).

2) Administration method: Two Boluoke capsules were taken orally half an hour before meal each time, three times a day for 21 days. During this period, other drugs that can dilate cerebral vessels, affect blood rheology and deoppilate the blood vessels were prohibited.

3) Appraisal of the clinical efficacy: The clinical neural deficit before and after the administration was scored by "The score criteria of clinical nervous function deficiency degree in patients with stroke" passed by the Second National Symposium on Cerebrovascular Diseases. Score 0-15 was defined mild; score 16-30 moderate and score 31-45 severe." Beijing score criteria of clinical efficacy in patients with stroke was adopted in the finalappraisal of efficacy (score declined more than 90% was defined basically recovered; score declined from 46% to 89% significantly improved, score declined from 18% to 45% improved; score declined less than 18% or increased less than 18% no changed and deficiency score increased more than 18% deteriorated). Some hospitals also studied the changes in fibrinogen and euglobulin lysis time. The blood was sampled at 8:00-8:30AM and the sampled site was ante-cubital vein.

4) Data processing: t-test and X2

test were applied with TheStatistic Software of The Capital Medical College Mathematics Faculty. All data were processed by IBM-PC.


(cont' d)

Results

The results indicated that the total effective rate was 88.21 %, and the significant response rate was 68.91 % (see Table 2). The results of those hospitals performed fibrinogen and euglobulin lysis time tests showed that fibrinogen and euglobulin lysis time were significantly reduced and the blood rheology was significantly changed after using Boluoke capsule (see Table 3 and Table 4). The adverse effects were very rare (see Table 5).

Discussion

Using earthworms as medicine was recorded in the Chinese traditional medicine masterpiece entitled Ben Cao Gang Mu (The Catalog of Herbal Medicine). It was deemed that the earthworm has the nature of warding off "wind" and unblocking the body's meridians and channels, and can be used to treat hemiplegia or pain syndromes due to "evil" blockages. In the 1983's international symposium of thrombosis and hemostasis, a Japanese scholar reported that the extract of earthworms had the effect of thrombolysis and denominated it "Lumbrokinase". In 1986, Korea Drugs Administration Bureau approved the manufacture of Lumbrokinase. Since then this enzyme has come into the market of many countries abroad and has been applied extensively. The trade name of Lumbrokinase in Hong Kong is "the Hart of Dragon". Boluoke capsule is a multi-component enzyme preparation and has the analogous component of t-PA, which can lyse the thrombi in vivo. There are at least two mechanisms: one is that Boluoke has the effect of plaminogen activator and the other is that it has special affinity to fibrin and thus degrades fibrin rapidly.

Because Boluoke capsule is an enzyme preparation, it is best taken half an hour before meals in order to reduce the effect of gastric acid. The low gastric volume and gastric acid enables the enzyme to be transported to intestinal tract as soon as possible.

In recent years, with further study on bioengineering technology, how large molecules such as protein penetrates the biomembrane has been extensively investigated. Fifteen years ago, Dr. Adiwl in US suggested that protein could be absorbed in vivo as peptides. In 1989, a Japanese professor verified this point by experiment. Many studies have found the general phenomena of trans-membrane transportation of protein and have elucidated theoretically that protein and enzyme can be transported transmembranely to all parts of the body and exert their normal physiological functions. Biophysics Institute of Chinese

Academy of Sciences has made animal models to verify the thrombolysis effect of Boluoke. 125j_ Labelled fibrinogen was intermingled when the thrombi were formed on the rabbit model for acute pulmonary artery embolism. Then Boluoke was given via duodenum and radioactive intensity was measured at the time of half-an hour, 1 hour, 2 hours and 5 hours post-administration. The result demonstrated that the radioactivity significantly increased 3 hours and 5 hours after administration, which means the effect of thrombolysis was significant 4-8 hours after administration. A Japanese professor also reported that the replicated venous thrombi of Beagle dog were dissolved overtly after oral administration of Lumbrokinase.

General Information of the Patients Table 1

Hospital No. of Patients Age Condition of the Illness

Mild Moderate Severe

Dur. of Illness Bef.

Trtmt (# of Days)

Pretreatment

Score Total Male Female 1 Beijing Xuanwu Hospital 210 126 84 61.51 ± 2.54 110 85 15 2.32 ± 2.54 17.16 ± 8.23

2 No. 2 Affiliated Hospital of Jiangxi Medical College 171 112 59 63.3 ± 9.13 89 73 9 3.61 ± 3.17 16.51 ± 7.11

3 Jiangxi Peoples Hospital 130 87 43 64.35 ± 9.83 105 20 5 26.72 ± 78.22 10.42 ± 7.50

4 Zhongshan Affiliated Hospital of Shanghai Medical College 50 27 23 66.49 ± 8.49 24 23 3 31.35 ± 37.59 15.51 ± 7.42

5 Affiliated Hospital of Tianjin Medical College

33 26 7 61.73 ± 9.38 10 17 6 26.72 ± 78.22 19.15 ± 11.36

6 Affiliated Hospital of West China Medical College 60 40 20 60.14 ± 7.78 28 27 5 6.10 ± 6.70 18.20 ± 9.15

7 Nanjing Brain Hospital 43 29 14 63.4 ± 8.78 33 7 3 13.74 ± 21.27 8.84 ± 9.75 8 Sichuan Peoples Hospital 50 39 11 66.2 ± 9.99 26 21 3 9.74 ± 23.76 16.2 ± 8.86

9 Affiliated Hospital of Chongqing Medical College 42 25 17 63.91 ± 9.29 29 12 1 6.21 ± 6.60 10.38 ± 8.77

10 No. 731 Hospital of Space Ministry 183 124 59 58.3 ± 9.52 120 44 19 7.23 ± 28.72 15.73 ± 8.89 11 No. 1 Handan Hospital 128 77 51 59.41 ± 8.67 95 31 2 2.72 ± 3.03 12.16 ± 7.35

12 Affiliated Hospital of Handan Second Medical School 97 53 44 62.00 ± 6.50 48 48 1 1.83 ± 2.17 16.67 ± 7.77

13 Handan Central Hospital 83 55 28 59.10 ± 8.26 46 33 4 3.33 ± 5.94 16.23 ± 8.14

14 Central Hospital of Handan Mineral Bureau 48 30 18 64.15 ± 5.28 28 20 0 1.50 ± 1.01 15.25 ± 7.13

15 An'yang Peoples Hospital 132 87 45 50.26 ± 8.07 80 42 10 2.97 ± 3.77 15.04 ± 9.87 16 Tangshan Workers' Hospital 100 64 36 61.31 ± 8.04 68 26 6 8.92 ± 10.15 13.54 ± 8.97

Total 1560 1001 559 62.38 ± 8.13 939 529 92 9.93 ± 14.20 13.59 ± 8.20

Clinical Response to Boluoke® (1560 cases total) Table 2

Hos itals p # of Patients Basically

Recovered Significantly

Improved Improved No Change Significant Response Rate

Total Effective Rate

1 Beijing Xuanwu Hospital 210 28 (13.33%) 98 (46.67%) 44 (20.95%) 40 (19.05%) 126 (60%) 170 (80.95%)

2 No. 2 Affiliated Hospital of Jiangxi Medical College 171 64 (37.43%) 59 (34.50%) 37 (21.64%) 11 (6.43) 123 (71.93%) 160 (93.57%)

3 Jiangxi People's Hospital 130 17 (13.08%) 51 (39.23%) 42 (32.31%) 20 (15.38%) 68 (52.31%) 110 (84.62%)

4 Zhongshan Affiliated Hospital of Shanghai Medical College 50 2 (4.00%) 16 (32%) 25 (50%) 7 (14%) 18 (36%) 43 (86%)

5 Affiliated Hospital of Tianjin Medical College 33 4 (12.12%) 14 (42.42%) 10 (30.3%) 5 (15.15%) 18 (54.55%) 28 (84.85%)

6 Affiliated Hospital of W. China Medical College 60 8 (13.33%) 25 (41.67%) 15 (25%) 12 (20%) 33 (55%) 48 (80%) 7 Nanjing Brain Hospital 43 21 (48.84%) 12 (27.91%) 4 (9.3%) 6 (13.95%) 33 (76.74%) 37 (86.05%) 8 Sichuan People's Hospital 50 4 (8%) 34 (68%) 9 (18%) 3 (6%) 38 (76%) 47 (94%)

9 Affiliated Hospital of Chongqing Medical College 42 18 (42.86%) 13 (30.95%) 4 (9.52%) 7 (16.67%) 31 (73.81%) 35 (83.33%) io No. 731 Hospital of Space Ministry 183 87 (47.54%) 76 (41.53%) 7 (3.83%) 13 (7.1%) 163 (89.07%) 170 (92.9%)

11 No. 1 Handan Hospital 128 19 (14.84%) 73 (57.03%) 20 (15.63%) 16 (12.5%) 92 (71.88%) 112 (87.5%)

12 Affiliated Hospital of Handan 2'd Medical School 97 22 (22.68%) 44 (45.36%) 23 (23.71%) 8 (8.25%) 67 (69.07%) 89 (91.75%)

13 Handan Central Hospital 83 26 (31.33%) 41 (49.40%) 8 (9.64%) 8 (9.64%) 67 (80.72%) 75 (90.36%) 14 Central Hospital of Handan Mineral Bureau 48 11 (22.92%) 19 (39.58%) 11 (22.92%) 7 (14.58%) 30 (62.5%) 41 (85.42%)

15 An'yang People's Hospital 132 29 (21.97%) 71 (53.79%) 21 (15.91%) 11 (8.33%) 100 (75.76%) 121 (91.67%)

16 Tangshan Workers' Hospital 100 21 (21%) 48 (48%) 21 (21%) 10 (10%) 68 (68%) 90 (90%) 17 Total 1560 381 (24.42%) 694 (44.49%) 301 (19.29%) 184 (11.79%) 1075 (68.91%) 1376 (88.21%)

**No. of patients that have deteriorated or died during the trial: 0% out of the 1560 cases**

Fibrinogen and Euglobulin Lysis Time Changes Table 3

Hospital

[§ P<0.05 , * P<0.01]

No. of

Pts

Fibrinogen Level (mg/di) Euglobulin Lysis Time (Minute)

Before Treatment After Treatment Before Treatment After Treatment

1 Beijing Xuanwu Hospital 210 355.86 +/- 55.98 294.77 +/- 45.15 * 196.89 +/- 70.24 153.40 +/- 48.00*

2 Jiangxi People's Hospital 130 414.39 +/- 159.15 356.50 +/- 103.88* 242.55 +/- 64.56 207.85 +/- 56.54*

3 Zhongshan Affiliated Hospital of Shanghai Medical College 50

352.14 +/- 307.31 208.38 +/- 138.14* N/A N/A

4 College Affiliated Hospital of Chongqing Medical

42 N/A N/A 176. 15 +/- 40.58 129.23 +/- 59.51§

5 No. 731 Hospital of Space Ministry 183 448.47 +/- 124.91 381.56 +/- 123.35* 194.16 +/- 71.80 156.83 +/- 55.36*

6 No. 1 Handan Hospital 128 304.03 +/- 39.55 295.78 +/- 34.35* 138.08 +/- 34.49 119.06 +/- 24.40*

7 Affiliated Hospital of Handan Second Medical School 97

391.54 +/- 70.66 311.71 +/- 61.34* 152.80 +/- 33.73 130.30 +/- 23.81*

8 Handan Central Hospital 83 376.87 +/- 56.32 323.48 +/- 58.91* 151.93 +/- 44.46 131.37 +/- 32.95*

9 Central Hospital of Handan Mineral Bureau 48 358.75 +/- 82.42 314.79 +/- 50.74* 165.81 +/- 33.99 130.46 +/- 24.53*

10 An'yang People's Hospital 132 390.26 +/- 54.95 334.25 +/- 39.67* 224.11 +/- 41.57 214.95 +/- 42.04*

11 Tangshan Workers' Hospital 100 330.05 +/- 83.28 269.88 +/- 58.89* N/A N/A

Blood Viscosity, Plasma Viscosity, Platelet Aggregation before and after Treatment Table 4

Hospital (*P<0.01, § P<0.05)

# of

Pts

Whole Blood Viscosity Plasma Viscosity

Pretrtmt After Trtmt

PAgT Pretrtmt After Trtmt Pretrtmt After Trtmt

1 Beijing Xuanwu Hospital 210 5.60 +/- 0.59 5.18 +/- 0.54* 1.83 +/- 0.12 1.72 +/- 0.09* 67.08 +/- 17.52 46.20 +/- 14.00 2 Zhongshan Affiliated Hospital of Shanghai

Medical College 50 7.28 +/- 1.64 6.48 +/- 1.47* 1.71 +/- 0.15 1.56 +/- 0.22* N/A N/A

3 Nanjing Brain Hospital 43 7.68 +/- 2.39 6.98 +/- 1.85 § 1.90 +/- 0.80 1.85 +/- 0.58 § N/A N/A 4 No. 721 Hospital of Space Ministry 183 9.35 +/- 2.51 8.30 +/- 1.72* 1.77 +/- 0.21 11.67 +/- 0.12* 61.14 +/- 18.12 51.70 +/- 18.89 5 No. 1 Handan Hospital 128 8.95 +/- 1.13 8.15 +/- 0.77* 1.81 +/- 0.11 1.70 +/- 0.08* 67.89 +/- 6.72 63.63 +/- 8.65

6 Central Hospital of Handan Mineral Bureau 48 6.77 +/- 0.87 6.20 +/- 0.72* 1.85 +/- 0.12 1.66 +/- 0.11* 71.25 +/- 11.33 65.06 +/- 4.75

Affiliated Hospital of Handan Secondary Medical School 97 7.02 +/- 1.27 6.02 +/- 1.21* 1.83 +/- 0.12 1.70 +/- 0.10* 70.01 +/- 7.00 64.98 +/- 5.78

8 Handan Central Hospital 83 7.80 +/- 1.65 6.92 +/- 1.45* 1.87 +/- 0.12 1.72 +/- 0.13* 67.11 +/- 9.59 61.28 +/- 5.2 9 An'yang People's Hospital 132 8.27 +/- 1.73 8.19 +/- 1.26* 1.70 +/- 0.11 1.57 +/- 0.15* 57.49 +/- 22.18 47.58 +/- 22.1 # Tangshan Worked Hospital 100 4.42 +/- 0.56 4.03 +/- 0.41* 1.69 +/- 0.14 1.00 +/- 0.09* 73.08 +/- 13.39 60.10 +/- 10.

Symptoms No. of Cases Percentage Skin Itching 9 0.58%

Skin Rash 3 0.19%

Nausea 18 1.15%

Out of 1,560 cases: Total Cases of Adverse Reactions

30 1.92%

Adverse Reaction Rates (out of 1560 cases) Table 5

BOLUOKE - IPlanet Health · 2015-12-19 · known allergy to lumbrokinase, earthworms or corn The above statements have not been evaluated by the FDA. This product is not intended

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