US008852.

578B2

(12) United States Patent (10) Patent No.: US 8,852,578 B2

Picard et al. (45) Date of Patent: Oct. 7, 2014

(54) COMBINATION THERAPY FOR (56) References Cited

TREATMENT OF PATIENTS WITH
NEUROLOGICAL DISORDERS AND U.S. PATENT DOCUMENTS
CEREBRALINEARCTION
5,942.233 A * 8/1999 Chang ........................... 424,758
2003/0211178 A1 11/2003 Xia
(75) Inventors: David Picard, Singapore (SG); Xuemin 2007/0059386 A1 3/2007 Lee ............................... 424,725
Shi, Tainjin (CN); Michael Lazdunski,
Nice (FR) FOREIGN PATENT DOCUMENTS
(73) Assignee: Moleac Pte. Ltd., Singapore (SG) CN 1435242 A 8, 2003
CN 1485.077 A 3, 2004
(*) Notice: Subject to any disclaimer, the term of this CN 1726966 A 2, 2006
patent is extended or adjusted under 35 CN 1973,865 A 6, 2007
CN 10.1088532 A 12/2007
U.S.C. 154(b) by 421 days. CN 101 194939 A 6, 2008
CN 101.244206 A 8, 2008
(21) Appl. No.: 13/128,529 WO WO-2007, 106049 A1 9, 2007

(22) PCT Filed: Nov. 10, 2009 OTHER PUBLICATIONS

(86). PCT No.: PCT/SG2O09/OOO416 Johnston, "Vinegar: medicinal uses and antiglycemic effect.”
Medscape General Medicine 8(2):61-72, 2006.*
S371 (c)(1), International Search Report for PCT/SG2009/000416 dated Jan. 19,
(2), (4) Date: Jul. 1, 2011 2010.
Gan, R. et al., “Danqi Piantan Jiaonang Does Not Modify
(87) PCT Pub. No.: WO2010/053456 Hemostasis, Hematology, and Biochemistry in Normal Subjects and
PCT Pub. Date: May 14, 2010 Stroke Patients.” Cerebrovascular Diease 25:450-456 (2008).
Hua Chiang Siow, C., “Neuroaid in Stroke Recovery.” European
(65) Prior Publication Data Neurology 60:264-266 (2008).
International Preliminary Report on Patentability for PCT/SG2009/
US 2011/025O183 A1 Oct. 13, 2011 000416 dated Jan. 18, 2011.
(30) Foreign Application Priority Data * cited by examiner
Nov. 10, 2008 (SG) ............................... 2008O8340-4 Primary Examiner — Rosanne Kosson
(74) Attorney, Agent, or Firm — Marshall, Gerstein & Borun
(51) Int. Cl. LLP
A6 IK36/00 (2006.01)
A6 IK35/12 (2006.01) (57) ABSTRACT
(52) U.S. Cl. The present invention provides compositions and methods for
USPC ........................... 424/93.7; 424/725; 424/520 treating stroke patients using TCM and a Western medica
(58) Field of Classification Search ment used for the treatment of stroke patients.
USPC ......................................... 424/93.7, 725, 520
See application file for complete search history. 19 Claims, 3 Drawing Sheets

Neurological deficit improvement on DTER score

before and after treatment

SEVERE
COMMON
MD
LOW

Before NeurOAid(6) After NeuroAid(E) Before BNJ After BN

U.S. Patent Oct. 7, 2014 Sheet 1 of 3 US 8,852,578 B2

DTER SC Ore S before and after treatments

SEVERE
OCOMMON
DMILD
DLOW
Before After
B NJ BN

Fig 1(a)

Functional Outcomes before and after treatment

&x---p-

Before NeuroAid(B) After NeuroAid(8) Before BNJ After BNJ

Fig 1(b)
U.S. Patent Oct. 7, 2014 Sheet 2 of 3 US 8,852,578 B2

Neurological deficit improvement on DTER score

before and after treatment

SEVERE
COMMON
:

Before NeuroAid6). After NeuroAid(6) Before BNJ After BNJ

Fig 2(a)

Comprehensive functions/Functional Outcomes

before and after treatments

Before NeuroAid B. After NeuroAid(B. Before BNJ

Fig 2(b)
U.S. Patent Oct. 7, 2014 Sheet 3 of 3 US 8,852,578 B2

Systolic blood pressure

Aspirin +
NeuroAid'
- Aspirin
alone

Diastolic blood pressure

ASpirin +
-0-NeuroAid'
-a-Aspirin
alone

Fig 3(b)
 US 8,852,578 B2
1. 2
COMBINATION THERAPY FOR eases (for example, multiple Sclerosis), cerebral palsy, trau
TREATMENT OF PATIENTS WITH matic injuries to or tumors in the brain, spinal cord and
NEUROLOGICAL DISORDERS AND peripheral nerves.
CEREBRAL INFARCTION NeuroAid R is capable of use as a Western medicine or a
dietary supplement to provide nutrition to healthy individuals
FIELD OF THE INVENTION as well as patients afflicted with stroke or neurological disor
ders.
The present invention provides methods and compositions NeuroAid R is typically administered orally (per os) as
for the treatment of stroke and neurological disorders. The Such or by diluting the capsules in water or via a gastric tube,
methods and compositions of the present invention bring 10 3 times each day and 4 capsules each time for a 4-week course
together aspects of Traditional Chinese Medicine (TCM) and of treatment. The duration of treatment is typically 3
Western medicine. months/3 courses, adaptable with regard to the patient's con
dition.
BACKGROUND The use of TCM is, however, particularly challenging for
15 European clinicians because of the lack of guidelines, clinical
Stroke is a major cause of death and disability. Primary data and the small number of studies conducted under West
stroke prevention focuses on lifestyle modifications of risk ern guidelines. Potential interactions between TCM and
factors while secondary stroke prevention aims to reduce the Western medicine that may lead to adverse side effects are
overall risk of recurrence in persons who have had a stroke. also a major concern among both practitioners of TCM and
There is currently a lack of treatment methods for stroke Western medicine. Of particular concern are the increase or
recovery in Western medicine, these being frequently limited decrease in the effects of a bloodthinner such as Warfarin that
to the following options: may lead to either a bleeding episode or formation of a blood
a) Intra-arterial thrombolysis with intravenous tissue plasmi clot, and the decrease in the effect of a blood pressure medi
nogen activator (rt-PA), which is applicable only to 3 to 5% cation that may lead to high blood pressure and a stroke. For
of stroke patients (as it has to be applied only for acute 25 example, the potential interaction of the Chinese herb salvia
ischemic forms, only if patients do not present any contra with the Western drug Warfarin leading to excessive blood
indication, and only within 3 to 6 hours after the onset of thinning with bleeding has been well documented with con
symptoms), can restore blood perfusion and prevent neu firmatory laboratory studies. Non-steroidal anti-inflamma
rological and functional damage to Some degree; tory drugs (NSAIDS), in particular aspirin, also have the
b) administration of aspirin/other antiplateletsfor sometimes 30 potential to interact with Chinese herbs and increase bleeding
anticoagulant to most of the cerebral stroke patients for risks. For these, and other reasons, TCM is generally not used
secondary stroke prevention, which gives an improvement in conjunction with Western medicines.
effect of about 1% (The International Stroke Trial (IST) a As current treatment options do not address the needs of
randomized trial of aspirin, Subcutaneous heparin, both or difficult-to-treat patients with important stroke disabilities,
neither among 19435 patients with acute ischaemic stroke. 35 Such as hemiparalysis or aphasia, the present invention seeks
International Stroke Trial Collaborative Group. Lancet. to combine TCM with established agents for the treatment of
1997 May 31; 349(9065): 1569-81); stroke patients to present a new therapeutic treatment option
c) medications such as analgesics may be needed to control for stroke patients.
associated symptoms
d) rehabilitation includes physical therapy Such as physio 40 SUMMARY
therapy, massage, speech therapy, or occupational therapy.
One known TCM product is NeuroAid R. Clinical studies According to a first aspect, there is provided a composition
performed in China on compositions the same as NeuroAid R. comprising:
have shown that this natural product combination increases (A) at least one of the following components: DanCui, Hon
stroke patients neurological disability recovery and func 45 ghua, Taoren, and Deer Horn; and
tional outcomes with extremely few side effects or other (B) at least one of the following components: Radix Astragali
adverse effects. The composition of NeuroAidR has been root (Membranous Milkvetch root or Huang Qi), Radix et
approved by and registered with the State Food and Drug Rhizoma Salviae Miltiorrhizae root (Red Sage root or Dan
Administration (SFDA) and is administered for the treatment Shen), Radix Paeoniae Rubra root (Red Peony root or Chi
of cerebral infarct patients during their recovery at an early or 50 Shao), rhizome of Ligusticum Chuanxiong (Chuan Xiong),
late stage, and the indications in Traditional Chinese Medi Radix et Rhizoma Notoginseng (Sandi), Cortex moutan
cine are: to supplement qi and activate blood circulation. It is (Peony or Mudanpi), Wood of Odoriferous Rosewood
applied to treat those patients who are Suffering from (Jiang Xiang), dried body of Scorpion (Quan Xie), Radix
ischemic or hemorrhagic (for the latter, it is at present indi Polygalae root (Yuan Zhi), Grassleaf sweetflag rhizome
cated in late phase only) cerebral infarction of qi deficiency 55 (Shi Changpu), Leeches (Hirudo or Shuizhi), Ground
and blood Stasis with manifestations of hemiplegia, hemian Beetle (or Tu Bie Chong), Natural or Artificial Cow-bezoar
esthesia, wry mouth, aphasia (inarticulateness) and etc during (calculus Bovis artifactus or Rengong Niuhuang), Gambir
their channel and collateral convalescent period of ischemia plant stem with hooks (Ramulus uncariae cum uncis or
apoplexy, and its efficacy is Supported by clinical trial data. Gou Teng).
It may be useful for treating other types of stroke than 60 In one embodiment, there is provided a composition as
cerebro-vascular stroke Such as cardio-vascular disease (heart defined in the first aspect, wherein for (A), the composition
stroke mainly due to coronary artery stroke) as well as other comprises at least 2, 3 or 4 of DanCui, Honghua, Taoren, and
neurological disorders. Neurological disorders are disorders Deer Horn. Optionally, for (B), the composition comprises at
that affect the central nervous system, the peripheral nervous least 4, 5, 6, 7, 8, 9, 10, 11, 12, 13 or 14 of the components as
system and the autonomic nervous system such as neurode 65 defined in the first aspect. In yet another embodiment, the
generative diseases (for example, Alzheimer's disease and composition further comprises an agent used in Western
Parkinson's disease), epilepsy, seizure, demyelinating dis medicine for the treatment of cerebral stroke, heart stroke,
 US 8,852,578 B2
3 4
neurodegenerative diseases, brain trauma, nervous system occurs when the flow of blood is blocked and oxygen cannot
trauma or conditions related to neuroplasticity. be delivered to the brain. Ischaemic stroke most commonly
Optionally, any one of (A) and (B) are present as separate occurs when the flow of blood is prevented by clotting
formulations. Furthermore, any one of the composition as (known as thrombosis of the artery) or by a detached clot
defined above and Western medicine may be present as sepa that lodges in an artery (referred to as an embolic stroke).
rate formulations. Haemorrhagic stroke results from rupture of an artery wall,
According to a second aspect, there is provided a compo and from blood leaking into the Surrounding brain. Haemor
sition as defined in the first aspect for use as a medicament. rhagic stroke, like ischemic stroke, causes the death of tissue
According to a third aspect, there is provided the use of the by depriving the brain of blood and oxygen, and results in a
composition as defined above, in the manufacture of a medi 10 number of neurological disabilities (motor, speech) as well as
cament for treating a patient having a condition selected from functional disabilities.
the group of cerebral stroke, heart stroke, neurodegenerative The term "stroke' refers to the sudden death of tissue cells
diseases, brain trauma, nervous system trauma and conditions due to a lack of oxygen when the blood flow is impaired by
related to neuroplasticity. blockage or rupture of an artery. Stroke is a vascular accident
According to a fourth aspect, there is provided a product 15 that can occur in the brain or in the cardiac system. The latter
comprising: condition is medically known as “myocardial infarction' and
a composition according to the invention; more commonly known as a “heart attack’. Because of the
an agent used in Western medicine for the treatment of stroke, similarity of both stroke mechanisms, it may be useful to use
for simultaneous, separate or sequential use in the treat NeuroAidR to help patients with a heart stroke recovering
ment of patient with a cerebral stroke, heart stroke, neuro better from their disability.
degenerative diseases, brain trauma, nervous system
trauma or conditions related to neuroplasticity; and DETAILED DESCRIPTION
optionally instructions for use of the product.
According to a fifth aspect, there is provided a kit treating The present invention provides a new combination treat
a patient for a cerebral stroke, heart stroke, neurodegenerative 25 ment for patients having one or more of the following condi
diseases, brain trauma, nervous system trauma or conditions tions: cerebral stroke, heart stroke, neurodegenerative dis
related to neuroplasticity, the kit comprising: eases, brain trauma, nervous system trauma or conditions
a composition according to the invention; and related to neuroplasticity.
instructions for administering said composition to a patient The present invention provides methods and compositions
The kit may further comprise an agent used in Western 30 for treating patients having a condition selected from the
medicine for the treatment of stroke, for simultaneous, sepa group consisting of cerebral stroke, heart stroke, neurodegen
rate or sequential use in the treatment of the patient. erative diseases, brain trauma, nervous system trauma and
According to a sixth aspect, there is provided a method of conditions related to neuroplasticity. The patients are admin
treating a patient having a condition selected from the group istered with:
of cerebral stroke, heart stroke, neurodegenerative diseases, 35 (A) at least one of DanCui, Honghua, Taoren, and Deer Horn;
brain trauma, nervous system trauma and conditions related and
to neuroplasticity, the method comprising administering to (B) at least one of a composition which comprises at least 1,
the patient a composition as define above. 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13 or 14 of the following
The method may further comprise administering an agent ingredients: Radix Astragali root (Membranous Milkvetch
used in Western medicine for the treatment of cerebral stroke, 40 root or Huang Qi), Radix et Rhizoma Salviae Miltiorrhizae
heart stroke, neurodegenerative diseases, brain trauma, ner root (Red Sage root or Dan Shen), Radix Paeoniae Rubra
Vous system trauma or conditions related to neuroplasticity. root (Red Peony root or Chi Shao), rhizome of Ligusticum
Chuanxiong (Chuan Xiong), Radix et Rhizoma Notogin
GLOSSARY OF TERMS seng (Sanqi), Cortex moutan (Peony or Mudanpi), Wood of
45 Odoriferous Rosewood (Jiang Xiang), dried body of Scor
This section is intended to provide guidance on the inter pion (Quan Xie), Radix Polygalae root (Yuan Zhi), Grass
pretation of the words and phrases set forth below (and where leaf sweetflag rhizome (Shi Changpu), Leeches (Hirudo or
appropriate grammatical variants thereof). Shuizhi), Ground Beetle (or Tu Bie Chong), Natural or
As used herein, the term “about as used in relation to a Artificial Cow-bezoar (calculus Bovis artifactus or Ren
numerical value means, for example, 50% or +30% of the 50 gong Niuhuang), Gambirplant stem with hooks (Ramulus
numerical value, preferably +20%, more preferably +10%, uncariae cum uncis or Gou Teng); and
more preferably still +5%, and most preferably +1%. Where (i) an agent used in Western medicine for the treatment of
necessary, the word “about may be omitted from the defini stroke.
tion of the invention. This invention is based on the discovery that NeuroAidR is
The term “comprising means “including or “consisting. 55 not only efficacious in treating stroke patients but that it may
Thus, for example, a composition "comprising X may con be safely used in combination with treatments commonly
sist exclusively of X or may include one or more additional used in Western medicine for treating stroke patients without
components. adverse side effects such as haemostasis disorders or high
The term “treatment includes any and all uses which blood pressure as described above.
remedy a disease state or symptoms, prevent the establish 60 As chronic forms of neurodegeneration Such as Alzhe
ment of disease, or otherwise prevent, hinder, retard, or imer's and Parkinson's disease (associated with cognitive
reverse the progression of disease or other undesirable symp dysfunction) share common mechanisms of neuronal death
toms in any way whatsoever. Hence, “treatment includes with acute forms of neurodegeneration which accompanies
prophylactic and therapeutic treatment. stroke, head trauma, cardiac arrest and Subarachnoid hemor
By “a cerebral stroke patient' we include a patient who has 65 rhage, NeuroAidR) may be useful to improve or treat these
Suffered an ischemic or haemorrhagic cerebral stroke. A cere brain disorders as it is believed to have potential activity on
bral stroke is a sudden and permanent death of brain cells that neuroprotection or on plasticity. Neuroplasticity (also
 US 8,852,578 B2
5 6
referred to as brain plasticity or cortical plasticity) refers to avoidance of doubt, by “an agent used in Western medicine'.
changes that occur in the organization of the brain and its we do not include medicaments used in TCM medicaments or
circuits of neurons, in particular changes that occur to the the like.
location of specific information processing functions. This Examples of Suitable agents include antiplatelets, antico
process Supports the learning of new functions as the result of 5 agulants, neuroprotectants, compounds for treating stroke
experience during development as mature animals and the and compounds for activating potassium channels TREK-1.
creation of new information with healthy neurons by-passing One, two, three, four, five or more of such further agents may,
damaged neurons affected by trauma or a medical condition. for instance, be used in combination with NeuroAid R. Thus,
Hence, in addition to treating cerebral stroke patients it is the following are envisaged as Suitable examples for use as
envisaged that NeuroAidR) or a NeuroAid R-like composition 10 combination partner (ii): an antiplatelet; an anticoagulant; a
(e.g. a composition according to (i) above), optionally in neuroprotectant; a compound for treating stroke; a compound
combination with available agents used in Western medicine, for activating potassium channels TREK-1; an antiplatelet in
may be useful for the treatment of such neurodegenerative combination with an anticoagulant; an antiplatelet in combi
diseases. Typically, the Western medicine used in combina nation with a neuroprotectant; an antiplatelet in combination
tion with NeuroAidR) or a NeuroAid R-like composition is 15 with a compound for treating stroke; an antiplatelet in com
one that targets a different mechanism from NeuroAidR) or a bination with a compound for activating potassium channels
NeuroAid R-like composition. For example, the Western TREK-1; an anticoagulant in combination with a neuropro
medicine may be antiplatelets and anticoagulants typically tectant; an anticoagulant in combination with a compound for
used in secondary stroke prevention, and neuroprotectants treating stroke; an anticoagulant in combination with a com
typically used in improving recovery potential in the acute 20 pound for activating potassium channels TREK-1; a neuro
phase of stroke via mechanisms described below. protectant in combination with a compound for treating
NeuroAidR and Similar Compositions stroke; a neuroprotectant in combination with a compound
The ingredients set forth in (i) above may be present in the for activating potassium channels TREK-1; a compound for
composition in a relatively crude form (e.g. unprocessed or treating stroke in combination with a compound for activating
crushed herbs) or in a more refined form (e.g. purified 25 potassium channels TREK-1; an antiplatelet in combination
extracts). with an anticoagulantanda neuroprotectant; an antiplateletin
In one embodiment, NeuroAid R is used. NeuroAid R is a combination with an anticoagulant and a compound for treat
TCM product in capsule form comprising 9 herbal compo ing stroke; an antiplatelet in combination with an anticoagu
nents and 5 animal components. NeuroAid R comprises lant and a compound for activating potassium channels
Radix Astragali root (Membranous Milkvetch root or Huang 30 TREK-1; an antiplatelet in combination with a neuropro
Qi), Radix et Rhizoma Salviae Miltiorrhizae root (Red Sage tectant and a compound for treating stroke; an antiplatelet in
root or Dan Shen), Radix Paeoniae Rubra root (Red Peony combination with a neuroprotectant and a compound for acti
root or Chi Shao), rhizome of Ligusticum Chuanxiong Vating potassium channels TREK-1; an antiplatelet in com
(Chuan Xiong), Radix et Rhizoma Notoginseng (Sanqi), Cor bination with a compound for treating stroke and a compound
tex moutan (Peony or Mudanpi), Wood of Odoriferous Rose- 35 for activating potassium channels TREK-1; an anticoagulant
wood (Jiang Xiang), dried body of Scorpion (Quan Xie), in combination with a neuroprotectant and a compound for
Radix Polygalae root (Yuan Zhi), Grassleaf sweetflag rhi treating stroke; an anticoagulant in combination with a neu
Zome (Shi Changpu), Leeches (Hirudo or Shuizhi), Ground roprotectant and a compound for activating potassium chan
Beetle (or Tu Bie Chong), Natural or Artificial Cow-bezoar nels TREK-1; an anticoagulant in combination with a com
(calculus Bovis artifactus or Rengong Niuhuang), Gambir- 40 pound for treating stroke and a compound for activating
plant stem with hooks (Ramulus uncariae cum uncis or Gou potassium channels TREK-1; a neuroprotectant in combina
Teng). NeuroAidR) may further comprise any one or more of tion with a compound for treating stroke and a compound for
DanGui, Honghua, Taoren, or Deer Horn activating potassium channels TREK-1; an antiplatelet in
NeuroAid R, which may be registered under different combination with an anticoagulant, a neuroprotectant and a
names in different countries (e.g. in South Africa it is mar- 45 compound for treating stroke; an antiplatelet in combination
keted as Strocaid R or Danqi Piantan Jiaonang.R.) is manufac with an anticoagulant, a neuroprotectant and a compound for
tured by and available commercially in the People's Republic activating potassium channels TREK-1; an antiplatelet in
of China from Tianjin Shitian Pharmaceutical Group Co., Ltd combination with an anticoagulant, a compound for treating
(located in the Jianxin Industrial area, Wangwenzhuang town, stroke and a compound for activating potassium channels
Xiqing district, Tianjin City, China; Postal Code 300381). It is 50 TREK-1; an antiplatelet in combination with a neuropro
also available from Moleac Pte Ltd (formerly Molecular Acu tectant, a compound for treating stroke and a compound for
puncture Pte Ltd), the main licensee outside of the People's activating potassium channels TREK-1; an anticoagulant in
Republic of China (11 Biopolis Way, Helios #09-08 Sin combination with a neuroprotectant, a compound for treating
gapore 138667; Tel: 6564789430; Fax: 6564789435). stroke and a compound for activating potassium channels
For the avoidance of doubt, NeuroAid R not only includes 55 TREK-1; and an antiplatelet in combination with an antico
NeuroAid R in the form in which it is currently marketed but agulant, a neuroprotectant, a compound for treating stroke
also includes future formulations of NeuroAidR which may, and a compound for activating potassium channels TREK-1.
for example, be marketed by Tianjin Shitian Pharmaceutical Antiplatelet Agents
Group Co., Ltd or Moleac Pte Ltd. Such future formulations Antiplatelet agents are medications that block the forma
may, for example, vary in dosage amounts or the concentra- 60 tion of blood clots by preventing the clumping of platelets.
tion of its active ingredients etc. Examples of antiplatelet agents include without limitations:
Combination Therapy with NeuroAidR Aspirin, the thienopyridine derivatives such as ticlopodine
In the present invention, NeuroAidR) can be used in com (Ticlid) and clopidogrel (Plavix), the Phosphodiesterase III
bination with one or more agents used in Western medicine inhibitors such as Cilostazol (Pletal), Adenosine re-uptake
for the treatment of stroke. By “an agent used in Western 65 inhibitors such as Dipyridamole (Persantine or AggrenoX (in
medicine, we include any form of mainstream medicine or combination with aspirin)), and the glycoprotein IIb/IIIa
Western medicine, including dietary supplements. For the inhibitors such as Abciximab (ReoPro), Eptifibatide (Integri
 US 8,852,578 B2
7 8
lin) and Tirofiban (Aggrastat) and orally active RGD mimetic opener (BMS 204352, cromakalim, levcromakalim,
prodrugs such as Orbofiban, Sibrafiban, SR 121566, or Roxi aprikalim, pinacidil, diazoxide, nicorandil, minoxidil), pirac
fiban. etam, adenosine transport inhibitor (propentofylline), gan
These agents differ in the way in which they prevent plate gliosides GM (non NMDA antagonist), presynaptic
lets from clumping: for example, Aspirin (Cyclo-oxygenase glutamate release inhibitors, claZosentan, desmoteplase,
inhibitor) blocks thromboxan A-2 by inhibiting the enzyme viprinex (ancrod), tenecteplase (TNKase; Metalyse),
cyclo-oxygenase-1 (COX-1), the thienopyridine derivatives alteplase, cyclic nitrones, TWEAK (TNF-like weak inducer
(ADP inhibitors) block the adenosine diphosphate (ADP) of apoptosis) receptor, thrombolytica treatments (urokinase,
receptor on the Surface of platelets membrane, and glycopro streptokinase, t-PA/tissue plasminogen activator or recombi
tein IIB/IIa inhibitors prevent platelet aggregation by inhib 10
nant urokinase), anistreplase, riluzole, and disufenton Sodium
iting a different receptor at the surface of platelets (the attach (NXY 059), candesartan, AX-200 (G-CSF, Fligrastim), caf
ment of glycoprotein IIb/IIIa to its receptor is the final step in feinol (caffeine-ethanol), enecadin, microplasmin, Sonoly
all pathways that cause platelets aggregation).
One or more antiplatelet agents can be employed in the sis+tPA, V-10153. HTUPA, solulin, piclozotan, S-0139,
present invention, for instance, a combination of, for 15 S-18986, AEOL-10150, AL-208, KN-38–7271, phridoxal
example, 2 or 3 or more antiplatelet agents may be employed. 5-phosphate, Neu-2000KL, ONO-2231, PGX-100, RVX
Anticoagulant Therapies 208, SUN-N4057, SUN-N8075, TAT-NR2B9c, GLP-1-ex
Antiplatelet agents are part of anticoagulation therapies. pressing stem cell therapy, Msc-1 (SA-4503, AGY-94.806)),
There are two other groups of anticoagulant agents which NH-02D, S-0139 259, tissue protective cytokines (Lu
may also be employed in the present invention: AA24493), V10153270 (BB-10153, TAPgen), combined use
Inhibitors of clotting factor synthesis such as without limita of statins and other cholesterol lowering drugs, erythropoi
tion, vitamin Kantagonists like coumarins or indanedione etin, cerebrolysin and CDP-choline (cytidine-5'-diphospho
derivatives (Warfarin or Coumadin). choline).
Inhibitors of thrombin that include several products such as, Compounds for Activating Potassium Channels TREK-1
but not limited to Heparin (Standard Unfractionated Hep 25 Other suitable agents used in Western medicine include
arin (UFH), Low Molecular Weight Heparin (LMWH) compounds capable of activating the potassium channels
Such as Enoxaparin, Tinzaparin) or to recombinant forms TREK-1. The activation of TREK-1 has been found to have a
of hirudin such as desirudin and lepirudin (Refludan). neuroprotective effect. In addition, TREK-1 has been shown
Neuroprotectants to play a major role in neuroprotection against epilepsy and
Using various mechanisms, neuroprotectants are com 30
brain and spinal cord ischemia. Since NeuroAidR) does not
pounds that preserve neuronal tissue at risk of dying during activate potassium channels TREK 1, compounds that are
stroke and in the aftermath of stroke. Some neuroprotectant capable of activating the potassium channels TREK-1 may be
agents are sometimes used to treat human stroke patients and used in combination with NeuroAid R to enhance the neuro
include antioxidants (e.g. selenium, 30 vitamin E. Vitamin C, protective effect of NeuroAidR).
glutathione, cysteine, flavinoids, quinolines, enzymes with: 35
reducing activity, etc), N-methyl-D-aspartate Receptor One example of a compound that is capable of activating
Antagonists (Dextrorphan, Selfotel, Magnesium), Narcotic the potassium channels TREK-1 is Polyunsaturated fatty
Receptor antagonist (Nalmefene (Cervene), Ca-channel acids (PUFAs). Polyunsaturated fatty acids are fatty acids that
blockers, Na-channel modulators (Lubeluzole), Alpha-ami contain more than one double bond. PUFAs can be catego
nobutyric acid agonist (Clomethiazole), glutamate receptor 40 rized as methylene-Interrupted Polyenes or Conjugated fatty
modulators, serotonin receptor agonists (repinotan), phos acids.
pholipids, free-radical scavenger (Tirilazad, and NXY-059), Methylene-Interrupted Polyenes are fatty acids that have
astrocyte activation inhibitor (ONO 2506), monoclonal anti two or more cis double bonds that are separated from each
bodies such as anti-ICAM-1 (Enlimomab), Human anti-leu other by a single methylene group. The essential fatty acids
kocytic antibody, Hu23F2G, membrane stabilization agent 45 are all omega-3 and -6 methylene-interrupted fatty acids.
CDP-choline (Citicholine), Fibroblast growth factor (Fi Examples of Omega-3 fatty acids include without limitation:
blast), unsaturated- and polyunsaturated fatty acids, estro Alpha-linolenic acid (ALA), Stearidonic acid (STD), Eico
gens and selective estrogen receptor modulators (SEAMS), satrienoic acid (ETE), Eicosatetraenoic acid (ETA), Eicosa
progestins, thyroid hormone and thyroid hormone-mimick pentaenoic acid (EPA), Docosapentaenoic acid (DPA),
ing compounds, cyclosporin A and derivatives, thalidomide 50 Docosahexaenoic acid (DHA), Clupanodonic acid, Tetra
and derivatives, methylxanthines, Mono-Amine-Oxydase cosapentaenoic acid, Tetracosahexaenoic acid or Nisinic
inhibitors (IMAO), serotonin-, noradrenaline and dopamine acid. Examples of Omega-6 fatty acids include without limi
uptake blockers, dopamine Iagonists, L-DOPA, nicotine and tation: Linoleic acid (LIN), Gamma-linolenic acid (GLA),
derivatives, and NO synthase modulators. Eicosadienoic acid, Dihomo-gamma-linolenic acid (DGLA),
Compounds for Treating Stroke 55 Arachidonic acid (AA), Docosadienoic acid, Adrenic acid,
Compounds used to treat human stroke patients include Docosapentaenoic acid or Osbond acid. Omega-9 fatty acids
Calcium Channel Blockers (D-600, Diltiazem, Nitrendipine, are also methylene-Interrupted polyenes, and may be mono
Nimodipine, Nifedipine, Flunarizine, Fluspirilene, Israd saturated or polysaturated. Examples of Omega-9 fatty acids
ipine, Nicardipine, PY 108-068, Verapamil and Triapamil), include without limitation: Oleic acid, Eicosenoic acid, Mead
Calcium chelator (DP-b99), free radical scavengers (Ebselen, 60 acid, Erucic acid or Nervonic acid.
Tirilazad, NXY-059), GABA receptor agonists (Diazepam, Conjugated fatty acids are fatty acids that have two or more
Baclofen), AMPA agonists (ZK 200775/MPQX), competi conjugated double bonds. Examples of Conjugated fatty
tive NMDA antagonists (aptiganel/cerestat, CP 101,606, dex acids include without limitation: Remenic acid, C.-Calendic
trophan, MK 801/dizocilpine, remacemide), glycine site acid, B-Calendic acid, Jacaric acid, C.-Eleostearic acid,
antagonists (GV 150526, ACEA 1021), polyamine site 65 B-Eleostearic acid, Catalpic acid, Punicic acid, Rumelenic
antagonists (eliprodil), growth factors (bFGF), Sodium chan acid, C-Parinaric acid, B-Parinaric acid, Bosseopentaenoic
nel blockers (fosphenytoin, 619C89), potassium channel acid.
 US 8,852,578 B2
9 10
Some other PUFAs which are not categorized as methyl highly contra-indicated and discontinued immediately if they
ene-Interrupted Polyenes or Conjugated fatty include without were part of patient's usual treatment. Protamine and Vitamin
limitation: Pinolenic acid and podocarpic acid. K may be given to reduce bleeding in patients with antico
Other compounds that may be capable of activating the agulant-induced bleeding.
potassium channels TREK-1 include the drug Riluzole (Ri 5 In addition to combination partners (i) and (ii), other com
lutekR), Lysophospholids (LPLs), Caffeic Acid esters and pounds may be administered, for example, proton pump
Xenon. These compounds may also be used in combination inhibitors such as Nexium, Protonix and Aciphex. Daily
with NeuroAidR to enhance the neuroprotective effect of doses of proton pump inhibitors are typically administered to
NeuroAid(R). reduce the risk of ulcer development and bleeding in patients
Modes of Administration 10 under long-term low-dose aspirin or antiplatelet therapy.
The combination partners (i) and (ii) may be present in a
single formulation or may be present as separate formula BRIEF DESCRIPTION OF DRAWINGS
tions. In one embodiment there may be a synergistic effect. As
mentioned above, combination partner (ii) may comprise FIG. 1 Phase II clinical trial results on use of NeuroAid R.
more than one agent, for example, two antiplatelet agents, or 15 versus BNJ (a) Neurological deficit improvement (DTER
an antiplatelet agent and a neuroprotectant may be used. scores) before and after treatment with NeuroAidR) or BNJ
The combination partners (i) and (ii) may be administered (b) Functional outcomes before and after treatment with Neu
to the patient at the same time (e.g. simultaneously) or at roAidR) or BNJ.
different times (e.g. sequentially) and over different periods FIG. 2 Phase III clinical trial results on use of NeuroAid R.
of time, which may be separate from one another or overlap versus BNJ (a) Neurological deficit improvement (DTER
ping. The combination partners (i) and (ii) may be adminis scores) before and after treatment with NeuroAidR) or BNJ
tered in any order. (b) Functional outcomes before and after treatment with Neu
The combination partner (ii) utilized and the appropriate roAidR) or BNJ.
administration route and dose level will be known to those in FIG. 3 Clinical study results on the interaction between
the art or could be readily determined by one skilled in the art. 25 NeuroAidR and aspirin in healthy volunteers and its effect on
Typically, as is well known in the medical art, dosage regi blood pressure. (a) systolic blood pressure (b) diastolic blood
mens may depend on various factors including the patients pressure.
size, body Surface area, age, the particular compound to be For FIGS. 1 and 2:
administered, sex, time and route of administration, general Level of severity of stroke (DTER scores)=LOW. MILD,
health, and other drugs being administered concurrently. 30
COMMON, SEVERE
While individual needs vary, determination of optimal ranges Functional outcomes: 0 pts=able to take care of oneself and
of effective amounts of each component is within the skill of speak freely; 2 pts able to live independently and to do
the art. The dosage would be similar to that administered Some simple work with Some incomplete functions; 4
when the agent is used without NeuroAid R. pts=able to walk and take care of oneself but must be
Dosage amounts for ticlopidine and for dipyridamole are 35 helped partially; 6 pts able to stand and take a step but
described in the Physicians’ Desk Reference, as are dosage must be taken care of at all times; 8 pts confined to bed.
amounts for other antiplatelet and neuroprotectant agents.
Dosage amounts of aspirin for the indicated effects are known EXAMPLES
to those skilled in the medical arts, and generally range from
about 20 mg to about 325 mg per day. For example, a formu 40 In the following examples, tables and drawings, the term
lation may contain about 20 mg. 30 mg, 80 mg, 160 mg, 250 “NeuroAidR' refers to a composition that is the same as
mg, 300 mg. 325 mg or 350 mg of aspirin. NeuroAid(R).
NeuroAid R may be administered orally as such, typically
with four 0.4 g capsules being taken 3 times a day. For patients Example 1
with Swallowing difficulties, capsules may be opened and 45
powder diluted in water that can be drunk as such or injected Phase II Trial: NeuroAid R. Versus Buchang
via a gastric tube. Hence, a daily dose of about 4.8 g is Naoxintong Jioanang (BNJ)
envisaged. In one embodiment, the patient’s daily dose of
NeuroAid R (or other composition according to (i) above) is A randomized, double-blinded, stratified, control design
about 2 g to 8 g; 3 g to 7 g; 4 g to 6 g; 4.25 g to 5.75 g; 4.5g 50 was adopted for the clinical trial on the efficacy of Neu
to 5.25 g; 4.5 g to 5 g; 4.6 g to 4.10g, or 4.7 g to 4.9g. A "daily roAid R in treating patients Suffering from apoplexy com
dose” can be a single tablet or capsule etc. or multiple tablets pared to BNJ which is known for its effectiveness in treating
or capsules etc. to be taken on a given day. Suitably, the patients suffering from apoplexy (see Example 3). A total of
composition according to (i) is taken orally. 200 subjects were involved: 100 cases were treated with
In one embodiment, each course of NeuroAid R treatment 55 NeuroAidR while 100 cases were treated with BNJ (control).
lasts about 4 weeks. Typically 3 courses are administered, BNJ is produced and provided by Xianyang Buchang Phar
most commonly back to back. No therapeutic window is maceutical Co., Ltd. Four capsules of each drug were admin
required but additional courses can be added even after a few istered 3 times daily, with each course of treatment lasting 4
days of treatment cessation. Hence, in one embodiment, each weeks.
NeuroAid R treatment lasts about 12 weeks. In another 60 The evaluation criteria for neurological and functional
embodiment, the treatment course of NeuroAid R (or other recovery from apoplexy (DTER scoring diagnostic standard)
composition according to (i) above) is about 4 to 24 weeks; 7 and TCM symptom therapeutic effects (TCM diagnostic
to 16 weeks; 9 to 15 weeks; 10 to 14 weeks; or 11 to 13 weeks. symptom scoring standard) were assessed in accordance with
In instances of ischemic stroke, treatment with anti-platelet the Clinical Guiding Principles for the Treatment of Apo
drugs usually commences as soon as possible after onset of 65 plexy with New Chinese Herbs promulgated by the Ministry
stroke symptoms while in instances of haemorrhagic stroke, of Health of the PRC in 1993. Severity of symptoms in the
anticoagulation treatments such as Coumadin or Heparin, are DTER scoring standard was classified according to 4 levels
 US 8,852,578 B2
11 12
(SEVERE, COMMON, MILD and LOW) while functional injection (60 mL) and BNJ for 1 month. The BNJ-treated
outcomes were classified in points from 0 to 8. group showed significantly better results in neurological and
The data (FIG. 1) demonstrated NeuroAidR's clinical effi functional outcomes.
cacy and Superiority in improving patients’ neurological defi Citicoline (CDP-choline) is a key intermediary in the bio
cit and functional (autonomy/dependency post-stroke) out synthesis of phosphatidylcholine, an important component of
comes versus that of the control treatment BNJ. the neural cell membrane that stabilizes cells membranes and
inhibits the formation of cytotoxic free fatty acids. It has been
Example 2 shown to produce beneficial effects in both animal models
and clinical stroke trials. A significant difference between the
Phase III Trial: NeuroAid(R) Versus BNJ 10 groups, favoring citicoline treatment, was seen in terms of
functional outcome as measured by the Barthel Index and
A randomized, double-blinded, stratified, control design Rankin scale, neurologic evaluation as measured by the
was adopted. A total of 405 subjects were involved, where National Institutes of Health (NIH) stroke scale, and cogni
300 cases were treated NeuroAid(R) while 105 cases were tive function as measured by the MiniMental Status Exami
treated with the control drug BNJ produced and provided by
15 nation. As efficacy trials on Citicoline have been demon
Xianyang Buchang Pharmaceutical Co., Ltd. Four capsules strated to be superior to the placebo, it can be inferred from
of each drug were administered 3 times daily, with each the above two comparative studies that treatment with BNJ
course of treatment lasting 4 weeks. would likewise be superior to a placebo.
The evaluation criteria for neurological and functional Example 4
recovery from apoplexy (DTER scoring diagnostic standard)
and TCM symptom therapeutic effects (TCM diagnostic Safety Study: Phase IV Open-Label Study on Effect
symptom scoring standard) were assessed as in the Phase II of NeuroAid R on Hemostasis
trial.
The data (FIG. 2) demonstrated that NeuroAidR was supe 25 Blood samples of 30 healthy individuals were collected
rior to BNJ in improving the patients neurological deficit before they received NeuroAidR) and 2 and 8 hours after
particularly in helping patients recover from their hemi-pa NeuroAid R initiation (4 capsules). Five coagulation tests
ralysis. With regard to functional outcomes, even if Neu were performed on the blood samples.
roAidR failed to demonstrate any superiority to BNJ, both
30 TABLE 1
treatments had comparable effect and about 50% of the stroke
patients included in the study returned to a functionality Blood Sampling First (%) Second (%) Third (%)
dependent state after 4 weeks of treatment (comprehensive
function score similar to Rankin score used in the West equal Quick Prothrombine Time
to 0 or 2). 35 Average 12.75 12.71 12.92
No adverse effects of treatment with NeuroAidR) were Std Deviation O.68 0.72 0.67
observed in the Phase II or Phase III trials. The finding of a Activated Partial Prothrombine Time
broad therapeutic window (>14 days after stroke onset) in Average 37.64 37.15 38.66
both trials also gives a huge advantage over fibrinolytic Std Deviation 4.28 4.44 4.26
agents, which have to be given within the first 3 to 6 hours. 40 Fibrinogen
Average 2.98 3.05 3.06
Example 3 Std Deviation O.61 O.S9 0.57
Platelet Aggregation
Comparative Study on Efficacy of BNJVersus
Average 63.01 62.19 61.52
Citicoline and Aspirin 45 Std Deviation 15.50 12.08 13.37
D-Dimer
A randomized, double-blinded, stratified, control design Control Device 2.7 2.4 1.9
was adopted for the comparative study on the efficacy of BNJ
and Citicoline in treating patients with apoplexy. Citicoline is
the only putative neuro-protectant that has shown results in 50 The results confirmed that NeuroAidR has no effect on
Western randomized, double-blinded trials given within 24 hemostasis blood factors and thus does not increase bleeding
hours after symptom onset. Davalos et al. (Oral Citicoline in risks.
Acute Ischemic Stroke. An Individual Patient Data Pooling
Analysis of Clinical Trials (2002) Stroke 33:2850) documents Example 5
the ability of Citicoline to improve complete recovery at 3 55
months. Safety Study: Phase IV Open-Label Study on
A first comparative study performed in the PRC involved Potential Interaction between NeuroAid R and
150 subjects treated with Citicoline (0.5 g IV) for 15 days in Acetylsalicylic Acid (Aspirin) in Healthy Volunteers
combination with a TCM (Xueshuantong) and 160 subjects and its Effect on Hemostasis and Blood Pressure
treated with Citicoline (0.5 g IV) for 15 days in combination 60
with Xueshuantong and BNJ. The latter group treated with An open randomized one-day study was conducted on 11
BNJ showed improvements in scores on a neuro-functional healthy volunteers receiving NeuroAid R treatment (12 cap
defects scale, plasma viscosity level and cholesterol level. sules per day) and aspirin (ASA 300 mg per day) from day 1
A second comparative study compared 30 subjects treated to day 5, and 11 healthy Volunteers receiving aspirin alone
with aspirin (150 mg), Citicoline (0.75 g IV) and salvia mil 65 (300 mg per day) from day 1 to day 5. Blood samples of the
tiorrhizae injection (60 mL), with 30 subjects treated with subjects were collected before they received treatment and 2
aspirin (150 mg), Citicoline (0.75 g IV), salvia miltiorrhizae and 8 hours after treatment initiation. The blood samples were
 US 8,852,578 B2
13 14
analyzed with the 5 coagulation tests, namely quick pro Example 6
thrombine time, activated partial prothrombine time, fibrino
gen dosage, platelet aggregation and D-dimer, to identify the Safety Study: Pilot Open-Label Study on Potential
coagulation mechanisms triggered by potential interaction Interaction between NeuroAidR) and Western Drugs
between NeuroAidR and aspirin. Blood pressure of the 22 5 in Ischemic Stroke Patients and its Effect on
Subjects was also measured before treatment, and 2, 4, 6 and Hemostasis and Blood Pressure
8 hours after treatment initiation.
Results of this trial show that NeuroAid R can be safely A pilot open-label study was conducted on 10 ischemic
associated with aspirin and does not have any interaction with 10
stroke patients within the first week of the onset of stroke
aspirin regarding blood coagulation (Table 2) and blood pres (early phase). The test patients were selected based on the
sure (FIG.3). following criteria:
a) aged above 18 years old;
TABLE 2 b) have had a cerebral infarction with compatible imaging on
15 Computed Tomography (CT) scan or Magnetic Resonance
Mean Imaging (MRI); and
Quick Prothrombine Time (Seconds) c) the time window between stroke onset and the open-label
study was less than one week,
Aspirin alone Conversely, patients that were excluded from the study
T=O 12.9 include:
T + 2 hours 12.6 a) female patients who were pregnant, lactating or nursing:
T+ 8 hours
Aspirin + NeuroAid (R)
12.4 b) patients showing signs of intra-cerebral hemorrhage on
brain Computed Tomography (CT) scan or Magnetic Reso
T=O 12.7 nance Imaging (MRI):
T + 2 hours 12.6 25 c) patients having a history of easy bruising or blood coagu
T+ 8 hours 12.4 lation disorders;
Activated Partial Prothrombine Time (Seconds) d) patients receiving other Traditional Chinese Medicine than
Aspirin alone NeuroAidR;
e) patients who received thrombolysis:
T=O 38.8
T + 2 hours 39.0
30 f) patients who have used NeuroAidR) within a 3-month
T+ 8 hours 38.2 period prior to screening and enrolment in the open-label
Aspirin + NeuroAid (R) study.
Each patient received 4 capsules of NeuroAid R 3 times a
T=O 37.1
T + 2 hours 38.2
day for one month in addition to the other Western medicine
T+ 8 hours 37.2
35 the patient was receiving. The Western medicine includes
Fibrinogen (G/L) platelet aggregation inhibitors, nitrates, oral anti-hyperten
sive drugs, lipid regulating drugs, oral anti-diabetic, or anti
Aspirin alone convulsant drugs.
T=O 3.2 Safety assessment tests were performed on blood samples
T + 2 hours 3.2 40 collected from the test patients at three intervals: a) before
T+ 8 hours 3.0 NeuroAid R intake (to form the baseline); b) 1 week after
Aspirin + NeuroAid (R) NeuroAid R initiation; and c) 4 weeks after NeuroAid R ini
T=O 3.2 tiation. The blood samples were analyzed using the following
T + 2 hours 3.0 hematology and biochemical laboratory analytical tests:
T+ 8 hours
Platelet Aggregation (Percentage)
2.9 45 a) Prothrombine Time (PT):
b) Activated Partial Prothrombine Time (APPT):
Aspirin alone c) fibrinogene dosage;
d) platelet aggregation;
T=O 62.82 e) D-dimer test;
T + 2 hours 62.87 50 f) blood cell count;
T+ 8 hours 48.01
Aspirin + NeuroAid (R) g) creatinine;
h) SGOT SGPT:
T=O 61.82 i) glycemia; and
T + 2 hours 58.28 j) CRP-C-reactive protein.
T+ 8 hours 46.98 55 The results of the open-label study (Table 3) demonstrated
D-Dimer (IG/L) that NeuroAidR) does not have any effect on renal and hepatic
Aspirin alone functions, glycemia, and C-Reactive-protein when used in
combination with Western medicine for managing risk fac
T=O O16 tors of secondary stroke. For example, in the five coagulation
T + 2 hours O16 60 tests (Prothrombine Time (PT), Activated Partial Prothrom
T+ 8 hours O16 bine Time (APPT), fibrinogene dosage, platelet aggregation,
Aspirin + NeuroAid (R) D-dimer test) performed on Patients 4 and 5 (both receiving
T=O O.13 NeuroAidR and four different types of Western medicine)
T + 2 hours O.09 (Table 3, columns 5 and 6), the test readings taken 1 week and
T+ 8 hours O.15 65 4 weeks after NeuroAid R initiation do not differ significantly
from the baseline readings taken before the start of Neu
roAid R intake.
 US 8,852,578 B2
15 16
The statistical analysis of the readings for the open-label tion (SD) values after 1 week and 4 weeks of NeuroAid R.
study (Table 4) also show that the readings from all 10 initiation are 1.22 and 1.11, respectively, which do not deviate
patients do not differ significantly from the mean values. For significantly from the baseline value of 1.13.
example, in the Prothrombine Time (PT) test, the Standard Hence, the results of the open-label study further demon
Deviation (SD) values after 1 week and 4 weeks of Neu- 5 strated that NeuroAidR) can be safely used in combination
roAidR initiation are 1.09 and 0.77, respectively, which do with Western medicine such as platelet aggregation inhibi
not deviate significantly from the baseline value of 0.88. tors, nitrates, oral anti-hypertensive drugs, lipid regulating
Similarly, in the fibrinogen dosage test, the Standard Devia- drugs, oral anti-diabetic, or anti-convulsant drugs.
TABLE 3
PATIENT 1. PATIENT 2 PATIENT 3 PATIENT 4 PATIENT 5

Gender F F F M F
Age 79 69 61 58 53
Day(s) from 3 4 1 2 7
stroke onset
On Aspirin? N Y N Y N
Other NeuroAid (R), NeuroAid (R), Neuro Aid (R), Neuro Aid (R), NeuroAid (R),
Treatment ISMN, aspirin, ISMN, aspirin, ISMN,
administered nimodipine nimodipine, metformin, nimodipine, nimodipine,
gemfibrozil glipizide glicazide, amlodipine,
indapamide KCI
NIHSS

Baseline 19 16 7 6
Week 17 O 8 4 4
Month 15 7 4 3
Rankin Score

Baseline 5 5 4 3
Week 5 4 2 1
Month 5 4 1 1
PT (secs)
Baseline 14.3 13.0 11.6 12.8 13.8
Week 12.7 11.6 11.1 12.4 14.4
Month 13.7 12.0 11.5 11.8 13.6
APTT (secs)
Baseline 27.1 32.1 29.0 33.5 33.1
Week 31.8 29.5 28.6 31.5 38.2
Month 29.6 29.9 3O.S 32.2 28.5
Fibrinogen
dosage (g/L)
Baseline 5.25 4.12 S.34 3.01 5.77
Week 6.76 3.03 3.38 3.80 4.64
Month 3.36 4.24 3.75 6.71 4.75
Platelet
agregation
Baseline 61-62 S1.2O 77.21 74.40 60.47
Week 47.8O 68.83 70.22 38.98 56.41
Month 60.82 71.92 33.11 37.30 42.59
D-Dimer (ng/mL)
Baseline O34 O.08 O.65 O.22 O16
Week 18O O.15 0.97 O.18 O.17
Month 2.38 O.24 O.71 O.2O O.20
SGPT - ALT
(IU/L)
Baseline 17.3 14.6 18.0 9.4 11.9
Week 15.3 28.0 38.7 15.2 20.1
Month 19.9 19.2 12.3 19.7 13.8
SGPT - AST
(IU/L)
Baseline 2O6 44.0 16.9 12.4 12.5
Week 21.9 18.8 32.O 14.8 16.8
Month 23.4 21.4 13.8 16.0 14.1
Glycemia
(mmol/L)
Baseline 9.28 4.71 10.14 6.36 3.93
Week 849 4.17 7.34 5.36 4.07
Month 6.83 4SO 5.89 S.26 3.99
 US 8,852,578 B2
17 18
TABLE 3-continued
Creatinine
(mol/l)
Baseline 53 53 46 81 43
Week 57 68 S4 95 51
Month 44 55 51 87 47
Red-blood cells

Baseline 4.32 4.15 4.57 S.10 3.48

4.18 4.07 4.67 S-31 3.73
3.70 4...SO 4.71 474 3.76

92.7 96.5 83.6 88.0 89.1

92.5 97.7 83.3 88.8 89.9
92.7 96.5 83.6 86.7 89.1

9.0 5.9 9.4 8.8 3.8

9.4 5.9 8.1 83 3.7
S.1 6.0 1O.O 12.8 3.5

7.9 4.1 6.4 4.9 2.4

7.8 3.5 4.6 4.0 2.3
3.6 3.7 6.3 8.8 1.9
ymphocytes
Baseline 0.7 1.3 2.2 3.0 O.9
Week 1.1 1.7 2.8 3.4 1.O
Month 1.O 1.6 3.0 2.8 1.3
Monocytes
Baseline O.3 0.4 OS
Week O.3 O.3 O6
Month O.3 0.4 0.7
eosinophiles
Baseline O.1 O.1 O1 O.2
Week O.1 O.2 O2 O.1
Month O.1 O.1 O3 O.1
basophiles
Baseline O.O O.O OO
Week O.O O.O OO
Month O.O O.1 O1
hemoglobin (Hb)
in g/L
Baseline 138 140 134 163 109
Week 131 139 138 167 120
Month 119 148 138 142 116
hematocrit (Hct)
in 96

Baseline 40.1 40.1 38.2 44.9 31.0

Week 45.8 39.8 38.8 47.1 33.5
34.3 43.4 39.4 41.1 33.5

6.34 O.99 6.42 O49 O.96

11.10 O.48 O.64 0.44 O16
O.64 O40 O.65 5.44 O.74

PATIENT 6 PATIENT 7 PATIENT 8 PATIENT 9 PATIENT 10

Gender F F M M M
Age 60 64 85 48 69
Day(s) from 7 1 1 4 7
stroke onset
On Aspirin? N N Y Y Y
Other Neuro Aid (R), Neuro Aid (R), NeuroAid (R), NeuroAid (R), Neuro Aid (R),
Treatment digoxine, phenytoin aspirin, aspirin, aspirin,
administered isosorbide Sodium, ISMN ISMN, ISMN,
mononitide, nimodipine amlodipine amlodipine
nimodipine,
phenytoin
 US 8,852,578 B2
19 20
TABLE 3-continued
NIHSS

Baseline
Week 12 3 3 1
Month 10
Rankin Score

Baseline
Week 5 2 2
Month
PT (secs)
Baseline 11.3 13.3 13.9 12.1 12.6
Week 11.5 12.8 13.9 12.0 11.6
Month 11.2 11.9 12.2 12.4 12.6
APTT (secs)
Baseline 27.9 29.3 33.5 303 307
Week 30.8 34.3 33.2 32.1 29.7
Month 24.9 29.8 30.2 347 307
Fibrinogen
dosage (gL)
Baseline 6.92 2.52 4.02 3.29 3.60
Week 3.42 4.78 3.01 3.09 3.77
Month 6.18 3.11 3.27 3.69 3.67
Platelet
agregation
Baseline 53.3 57.83 69.55 66.29 53.90
Week 48.51 37.53 54.97 45.68 50.37
Month 63.37 51.27 51.93 45.98 41.07
D-Dimer (ng/mL)
Baseline O.20 O.30 O.10
Week O.22 O.30 O.18
Month O.22 O.10 O.22
SGPT - ALT
(IU/L)
Baseline 18.8 20.3 8.9 14.5 9.9
Week 17 28.9 8.1 21.4 11.5
12 33.4 9.7 21.1 10.8

18.3 12.5 23.2 17.4 12.8

30.2 16.7 13.1 17.6 11.1
22.7 15.4 15.9 13.8 11.3

6.34 4.38 4.60 4.51 S.O3

5.45 3.59 4.44 423 5.07
6.4 3.53 5.24 4.05 4.78

Baseline 72 100 90 86 83
Week 47 93 105 85 72
Month 52 89 88 82 68
Red-blood cells

Baseline 4.13 4.29 4.79 5.14 3.90

4.33 4.64 4.60 S.16 4.24
4.4 4.67 5.36 S-13 4SO

89.6 92.0 85.6 89.7 99.2

89.2 91.4 85.2 91.7 97.9
88.4 92.1 83.2 86.9 1OO.O

7.8 4.2 7.7 7.2 3.7

11.7 6.9 S.1 S.2 3.8
7.2 6.2 6.O 5.7 4.8
 US 8,852,578 B2
21 22
TABLE 3-continued
granulocytes
Baseline 5.3 2.8 S.O 5.7 2.8
Week 7.7 5.4 3.1 3.2 3.0
Month 3.9 4.7 3.6 3.7 3.6
ymphocytes
Baseline 1.3 1.O 1.6 13 0.7
Week 2.9 O.9 1.8 1.7 0.7
Month 2.8 1.3 2.1 1.6 1.O
Monocytes
Baseline O.S O.2 O2
Week O.6 O.3 O3
Month O.S O.2 0.4
eosinophiles
Baseline O.1 0.4 O.1
Week O.1 O.1 O.1
Month O.1 O.1 O.1
basophiles
Baseline O.1 O.1 O.1
Week O.O O.1 O.1
Month O.O O.1 O.1
hemoglobin (Hb)
in g/L
Baseline 141 130 137 162 132
Week 147 132 137 161 146
Month 153 133 162 157 158
hematocrit (Hct)
in 96

Baseline 39.5 37 41.O 46.1 38.7

Week 42.4 38.7 39.2 473 41.5
Month 43.0 38.9 44.6 44.6 45.0
CRP (mg/dL)
Baseline O.14 3.53 2.17 O.69 O.17
Week 3.73 2.1 O3S O.13 O.10
Month 1.10 1.88 O.10 O38 O.10

TABLE 4 TABLE 4-continued

40
SUMMARY MEAN SD SD SD SUMMARY MEAN SD SD SD

Gender F =S M=4 Fibrinogen dosage

Age 65.11 53.24 76.98 11.87 (gL)
Day(s) from stroke 3.33 O.94 5.73 240
Onset 45 Baseline 4.10 2.97 5.23 1.13
On Aspirin? Y=S N=4 Week 4.O3 2.81 5.25 122
Other Treatment Month 4.06 2.95 S.18 1.11
administered Platelet agregation
NIHSS
50 Baseline 63.61 54.67 72.55 8.94
Baseline 7.11 O.86 13.36 6.25 Week 52.31 40.68 63.94 11.63
Week 4.89 -0.17 9.95 S.O6 Month 48.44 36.31 60.58 12.14
Month 4.11 -0.37 8.60 4.48
D-Dimer (ng/mL)
Rankin score
Baseline O.25 O.08 O42 O.17
Baseline 3.11 1.57 4.65 1.54 55
Week O45 -0.12 1.02 0.57
Week 2.22 O.83 3.62 1.39
Month O49 -0.23 122 0.73
Month 1.78 O.21 3.34 1.56
PT (secs) SGPT - ALT (IU/L)

Baseline 13.04 12.17 13.92 O.88 Baseline 13.87 9.75 17.98 4.11
60 Week 20.80 11.13 30.47 9.67
Week 12.SO 11.41 13.59 1.09
Month 12.41 11.64 13.18 O.77 Month 17.77 10.48 25.06 7.29
APTT (secs) SGOT - AST (IU/L)
Baseline 30.96 28.70 33.21 2.26 Baseline 1914 9.03 29.26 10.11
Week 32.10 29.17 35.03 2.93 65 Week 18.09 12.00 24.18 6.09
Month 30.68 28.87 32.48 18O Month 16.12 12.26 19.99 3.87
 US 8,852,578 B2
24
TABLE 4-continued Cos-7 cells, cultured in Dulbecco's Modified Eagle
Medium (DMEM; Gibco, Invitrogen, San Diego, Calif.)
SUMMARY MEAN SD- SD- SD supplemented with 10% fetal bovine serum (Perbio, Thermo
Glycemia (mmol/L) Fisher Scientific Inc., Waltham, Mass.), 100 U/ml penicillin
and 100 g/ml streptomycin (Gibco, Invitrogen, San Diego,
Baseline 5.88 3.60 8.16 2.28 Calif.) in a humidified incubator with 5% CO, at 37°C., were
Week S.21 3.44 6.98 1.77
Month 4.91 3.81 6.02 1.10
used for electrophysiological procedures. Cells were trans
Creatinine (mol/l) fected using DEAE-Dextran (Sigma-Aldrich Corporation,
St-Louis, Mo.) with 0.5ug of TREK 1 or TREK 2 cloned in
Baseline
Week
70.56
75.56
48.97
55.81
92.14
95.30
21.58
19.75
10 pIRES-EGFP plasmid (Clontech Palo Alto, Calif.) express
Month 67.89 48.94 86.84 18.95
ing the green fluorescent protein.
Red-blood cells Recordings were realized 48 hours after transfection. For
electrophysiological recordings, the internal solution con
Baseline
Week
4.42
4.51
3.87
4.OO
4.96
S.O2
0.55
O.S1
tained 155 mM KC1, 3 mM MgCl, 5 mM EGTA and 10 mM
Month 4.56 4.01 S.11 0.55
15 HEPES (Sigma-Aldrich Corporation, St-Louis, Mo.) at pH
Mean RBC volume 7.2 with KOH and the external solution contained 150 mM
NaCl, 5 mM KC1, 3 mM MgCl, 1 mM CaCl and 10 mM
Baseline 90.71 85.73 95.70 4.99 HEPES at pH 7.4 with NaOH. Effects of NeuroAidR) on
Week 90.93 86.OO 95.86 4.93
Month 90.09 84.36 95.82 5.73 TREK currents were measured on whole-cell currents, elic
white-blood cells ited by voltage ramps (from -100 mV to +100 mV, 1.5 sec
onds in duration) before and during application of Neu
Baseline
Week
6.63
5.88
4.33
4.10
8.94
7.65
2.30
1.77
roAidR) (5 minutes). TREK1 and TREK 2 currents were not
Month 6.68 3.79 9.57 2.89 different before and after 5 minutes of NeuroAidR) applica
granulocytes tion, comparing with control cells transfected with EGFP
25 only.
Baseline
Week
4.67
4.10
2.83
2.43
6.51
5.77
1.84
1.67
The results show that NeuroAidR) does not have any effect
Month 4.43 2.43 6.44 2.01 on potassium channels TREK-1. Hence, the results suggest
ymphocytes that agents that activate potassium channels TREK-1 and
NeuroAidR have different mechanisms and different targets.
Baseline
Week
1.41
1.68
O.65
O.77
2.17
2.59
O.76
O.91
30 Accordingly, it may be beneficial to combine agents that
Month 1.74 1.01 2.48 O.74 activate potassium channels TREK-1 with NeuroAidR to
monocytes enhance the neuroprotective effect of NeuroAid R.
It will be understood that the invention has been described
Baseline
Week
O.31
O.29
O.15
O.15
O.47
O.43
O16
O.14
by way of example only and modifications may be made
Month O.34 O.19 O.SO O.15
35 whilst remaining within the scope and spirit of the invention.
eosinophiles
The invention claimed is:
Baseline O.11 O.OS O.17 O.O6 1. A product comprising:
Week O.13 O.08 O.18 O.OS
Month O.13 O.O6 O.20 O.O7 (a) a therapeutically effective amount of Chinese Angelica
basophiles 40 root (Radix angelicae Sinensis; Dan Gui); and
(b) therapeutically effective amounts of at least three of the
Baseline
Week
O.O3
O.O2
-0.02
-0.02
O.O8
O.O7
O.OS
O.04
following components: Membranous Milkvetch root
Month O.04 -0.01 O.10 O.OS
(Radix astragali root; Huang Qi); Red Sage root (Radix
hemoglobin (Hb) in et Rhizoma salviae miltiorrhizae; Dan Shen); Red Peony
45 root (Radix paeoniae rubra root; Chi Shao); rhizome of
Baseline 139.56 12341 155.70 16.15
Ligusticum chuanxiong (Chuan Xiong); Radix et Rhi
Week 142.89 128.42 157.36 1447
Zoma Notoginseng (Sanqi); Peony (Cortex moutan;
Month 143.67 126.96 160.38 16.71 Mudanpi); Wood of Odoriferous Rosewood (Jiang
hematocrit (Hct) in % Xiang); dried body of Scorpion (Quan Xie): Radix
Baseline 39.96 35.65 44.26 4.31
50 polygalae root (Yuan Zhi); Grassleaf Sweetflag rhizome
Week 41.71 37.19 46.23 4.52 (Shi Changpu); Leeches (Hirudo; Shuizhi); Ground
Month 40.99 36.58 45.40 4.41 Beetle (Tu Bie Chong); Natural or Artificial Cow-bezoar
CRP (mg/dL) (Calculus bovis artifactus; Rengong Niuhuang); and
Baseline 2.04 -0.49 4.57 2.53
Gambirplant stem with hooks (Ramulus uncariae cum
Week 1.90 -1.73 S.S4 3.63
55 uncis; Gou Teng).
Month 1.06 -0.61 2.73 1.67 2. The product of claim 1, further comprising:
(c) a therapeutically effective amount of an agent used in
Western medicine for the treatment of cerebral stroke,
Example 7 heart stroke, neurodegenerative diseases, brain trauma,
60 or nervous system trauma.
3. The product of claim 1, wherein (a) and (b) are present in
Effect of Neuro Aid R on Specific Potassium separate formulations.
Channels 4. The product of claim 1, further comprising:
(c) an agent used in Western medicine which is capable of
NeuroAidR powder was dissolved in distilled water at a 65 activating potassium channel TREK-1.
concentration of 1 Jug/ml. After 30 minutes infusion, the liq 5. The product of claim 4, wherein (a), (b), and (c) are
uid was filtered on a 0.22 um filter. present in separate formulations.
 US 8,852,578 B2
25 26
6. The product of claim 4, wherein the agent used in West 11. The product of claim 10, wherein the agent used in
ern medicine which is capable of activating potassium chan Western medicine which is capable of activating potassium
nel TREK-1 is a polyunsaturated fatty acid (PUFA), riluzole channel TREK-1 is a polyunsaturated fatty acid (PUFA).
(RilutekR), a lysophospholid (LPL), a caffeic acid ester, or riluzole (Rilutek.R.), a lysophospholid (LPL), a caffeic acid
XCO.
ester, or Xenon.
7. The product of claim 4, further comprising: 12. The product of claim 10, further comprising:
(d) instructions for administering the composition and the (d) instructions for administering the composition and the
agent to a patient. agent to a patient.
8. The product of claim 4, wherein (a), (b), and (c) are 13. The product of claim 10, wherein (a), (b), and (c) are
provided for simultaneous, separate, or sequential adminis 10
provided for simultaneous, separate, or sequential adminis
tration to a patient. tration to a patient.
9. The product of claim 1, wherein (a) and (b) are present in 14. A method for treating cerebral stroke, heart stroke, a
a single formulation. neurodegenerative disease, brain trauma, or nervous system
10. A product comprising:
(a) at least one of the following components, each compo 15 trauma in a patient, comprising administering the product of
nent present in a therapeutically effective amount: Chi claim 1 to the patient.
nese Angelica root (Radix angelicae sinensis; DanCui), 15. The method of claim 14, wherein the method further
Safflower (Carthamus tinctorius; Hong Hua), Peach comprises administering an agent used in Western medicine
seed (Prunus persica; Taoren), and Deer Horn; and which is capable of activating potassium channel TREK-1.
(b) at least one of the following components, each compo 16. The method of claim 15, wherein the agent used in
nent present in atherapeutically effective amount: Mem Western medicine is administered simultaneously, separately,
branous Milkvetch root (Radix astragali root; Huang or sequentially to the product.
Qi); Red Sage root (Radix et Rhizoma salviae miltior 17. A method for treating cerebral stroke, heart stroke, a
rhizae; Dan Shen); Red Peony root (Radix paeoniae neurodegenerative disease, brain trauma, or nervous system
rubra root; Chi Shao); rhizome of Ligusticum chuanx 25 trauma in a patient, comprising administering the product of
iong (Chuan Xiong); Radix et Rhizoma Notoginseng claim 6 to the patient.
(Sanqi); Peony (Cortex moutan; Mudanpi); Wood of 18. A method for treating cerebral stroke, heart stroke, a
Odoriferous Rosewood (Jiang Xiang); dried body of neurodegenerative disease, brain trauma, or nervous system
Scorpion (Quan Xie): Radix polygalae root (Yuan Zhi); trauma in a patient, comprising administering the product of
Grassleaf Sweetflag rhizome (Shi Changpu); Leeches 30
claim 2 to the patient.
(Hirudo; Shuizhi); Ground Beetle (Tu Bie Chong); 19. A method for treating cerebral stroke, heart stroke, a
Natural or Artificial Cow-bezoar (Calculus bovis arti neurodegenerative disease, brain trauma, or nervous system
factus; Rengong Niuhuang); and Gambirplant stem with trauma in a patient, comprising administering the product of
hooks (Ramulus uncariae cum uncis; Gou Teng); and claim 4 to the patient.
(c) an agent used in Western medicine which is capable of 35
activating potassium channel TREK-1. k k k k k