FORM 2
THE PATENTS ACT 1970 .
(39 of 1970)
AND
The Patents Rules, 2003
COMPLETE SPECIFICATION
(See section 10 and rulel3)
1. TITLE OF THE INVENTION:
"NOVEL PHARMACEUTICAL COMBINATION COMPRISING CITICOLINE AND FOLIC ACID"
2. APPLICANT:
(a) NAME: Lyka Labs Limited.
(b) NATIONALITY: Indian Company incorporated under the
Companies Act, 1956
(c)ADDRESS: 101, Shivshakti Industrial Estate, Andheri-Kurla Road,
Andheri (East), Mumbai - 400059, Maharashtra, India,
3. PREAMBLE TO THE DESCRIPTION:
The following specification particularly describes the invention and the manner in which it is to be performed:

Technical field:
This invention relates to novel pharmaceutical combination comprising Citicoline in association with folic acid in a solid oral dosage form, preferably in tablet form; useful for the treatment of neurodegenerative disorders. The inversion also relates to the process for preparation thereof.
Background and prior art:
Neurodegenerative disorders are characterized by progressive and irreversible loss of neurons from specific regions of the brain. Neurodegenerative disorders include Parkinson's disease, Huntington's disease, Alzheimer's disease and amyotrophic lateral sclerosis.
As a group, these disorders are relatively common and represent a substantial medical and social problem. They are primarily disorders of elderly, developing individuals who are neurologically normal.
At present the pharmacological therapy of neurodegenerative disorders is limited to symptomatic treatments that do not alter the course of the underlying disease. There is a need for more effective strategies to arrest or alter the course of the disease.
Cognitive functioning declines and prevalence and incidence of dementia rise with advancing age. Most dementia sufferers (about 65%) are the fastest growing segment of elderly populations in our country.
In a large ambulatory population of consecutive elderly subjects (60-plus years old, mean age 77.5 years), it was recently found that relative deficiency, besides being associated with Alzheimer's disease and vascular dementia, may also precede dementia onset. This finding goes along with the association of low serum folate with all types of dementia reported in the Canadian Study of Health and Aging and the strong negative correlation between folate and cortical atrophy on autopsy found in the Nun study, a correlation previously suggested also by a neurophysiological and neuroimaging in 16 patients with neurological and psychiatric disorders. Moreover, in a case-control study, Clarke et al. found that low folate and vitamin B12 and elevated homocysteine were associated with histologically confirmed Alzheimer's disease.

. Folate is an important factor for the normal development and functioning of the central nervous system at all ages. Iskandar and colleagues showed that the effect of folic acid supplementation is not restricted to the embryonic period but can also enhance growth, repair and recovery in the injured central nervous system of adult rats. By limiting the synthesis of methionine, folate deficiency can lead to both an accumulation of homocysteine and a decreased production of S-adenosylmethionine (SAM), a major methyl donor in a variety of methylation reactions important to the central nervous system. SAM concentration has been found to be severely decreased in the brain of Alzheimer disease patients. Folic acid deficiency and high homocysteine have been reported to promote cortical neurodegeneration. High dose Folic acid has been found to improve endothelial function independent of its homocysteine-lowering effect.
Alzheimer's disease is a neurodegenerative disease that destroys brain cells, resulting in memory loss and problems with thinking, behavior, lifelong hobbies and social life. It is a progressive and fatal disease, without a current cure, It affects both men and women usually 65 years and older. But an early onset form also exists. Alzheimer's is the seventh leading cause of death in United States; as many as 5 million Americans are living with the disease, according to the National Alzheimer's Association (Alz.org).
One of the characteristics of Alzheimer's disease is the deposition of senile plaques containing amyloid-beta (A-beta), a protein that promotes inflammation and oxidation. Natural supplementation is becoming a popular preventive method for Alzheimer's disease and other neurodegenerative diseases, by enhancing memory and cognitive functions.
High blood concentrations of Folic acid, a form of the water-soluble vitamin B9, are associated with positive cognitive function. In 2008 six-month, randomized, double blind, placebo-controlled study, 57 outpatients with probable Alzheimer's disease were treated concurrently with cholinesterase inhibitors (ChI) and either folic acid or a placebo. At the end of the six-months, there were no significant baseline differences or use of individual Chi between the two arms. A significant difference was seen in the change from baseline in combined IDAL and Social Behavior scores between arms versus the placebo, but there was no change in MMSE scores. Sixteen of 23 subjects receiving folic acid and seven out of 18 placebo subjects were classified as NICE responder (P=0.05).

Researchers concluded that response to ChI in patients with Alzheimer's may be improved by the use of folic acid.
A study published in The Lancet randomly assigned 818 participants 800μg/d folic acid or placebo for three years. Serum folate concentrations increased by 576 % and plasma total homocysteine concentrations decreased by 26% in participants taking folic acid compared with those taking placebo. The three-year change in memory, information processing speed and sensor meter speed were significantly better in the folic acid group than in the placebo group.
In an Italian study published in the Journal of the American College of Nutrition, 471 consenting subjects participating in the Monzino 80-plus study, a door-to-door population survey among very old subjects living in Northern Italy, were assessed for their serum vitamin B12 and folate concentrations. Cognitive and functional evaluations included MMSE (Mini Mental State Examination), IADL and Spontaneous Behavior Interview-basic Activities of Daily Living (SBI-bADL). The MMSE, IADL and SBI-bADL scores were all significantly correlated with the folate concentrations, while no significant associations were found with Vit B12 concentrations. Folate showed a highly significant, curvilinear association with both cognitive and functional scores (P<0.0001). Subjects in low and middle folate tertiles had significantly higher odds ratios for dementia. The findings suggested subclinical folate deficiency may represent a risk factor for the cognitive decline associated with aging that could contribute to Alzheimer's as well as other dementia development.
Citicoline, an intermediate in the generation of phosphatidylcholine from choline usually grouped within the Vitamin B complex to supports phospholipid production in the brain, has been used to treat neurodegenerative disorders. In a double blind, placebo-controlled, randomized clinical trial, 30 patients with mild to moderate senile dementia of the Alzheimer type were treated with 1,000 mg/d of citicoline (as Cognizin™, from Kyowa Hakko) (n=13) or placebo (n=I7) for 12 weeks. Compared to placebo, citicoline improved cognitive performance in Alzheimer's disease patients with apolipoprotein E (APOE); this improvement was more pronounced in patients with mild dementia. Citicoline also increased cerebral blood flow velocities in comparison with the placebo

(P<0.05) when transcranial Doppler recordings from both hemispheres were considered together, as well as diastolic velocity in the left middle cerebral artery (P0.05). Patients treated with citicoline showed an increase in the percentage of "brain bioelectrical activity of alpha (occipital electrodes) and theta type (left side electrodes), accompanied by a decrease in relative delta activity particularly marked in the left temporal lobe. Significant differences with respect to placebo (P0.05) were observed for theta activity in several fronto-parieto-temporal electrodes of the left hemisphere. Treatment with citicoline tended to reduce serum IL-1 beta levels, mainly after four weeks of administration, with no modified blood histamine content. In addition, neither adverse side effects nor alterations in biological and hematological parameters were induced by citicoline.
Combination of Citicoline with Folic Acid- a therapeutic need
The most striking feature of the combination is the requisite specificity of Citicoline and Folic acid as neuroprotective agents. Citicoline achieves therapeutic effect by its ability to increase the synthesis of phosphatidylcholine, the primary neuronal membrane component. Besides it enhances acetylcholine synthesis which ameliorate symptoms caused by loss of cholinergic nerves, Citicoline also influences the outcome of neurodegenerative diseases by its ability to reduce free fatty acid accumulation at the site of injury which prevents further damage. Folic acid improves memory and cognitive functions.
Citicoline and Folic acid may be administered for long term in elderly patients without any serious side effects.
Citicoline and Folic acid may be termed as an ideal combination for the management of neurodegenerative or ischaemic cerebral diseases, conditions in which patients suffer till death.
Therefore, the present inventors have invented novel pharmaceutical combination comprising therapeutically effective amount of Citicoline and water insoluble Folic acid along with pharmaceutically acceptable excipients.

Object of the Invention:
The main object of the invention is to provide novel pharmaceutical combination comprising therapeutically effective amounts of Citicoline in association with Folic acid useful for the treatment of neurodegenerative disorders, in solid oral dosage form along with pharmaceutically acceptable excipients such as diluents, binders, disintegrants, lubricants, glidants and the process for preparation thereof.
Summary of the Invention:
Accordingly, in one aspect the present invention describes solid oral dosage formulation comprising therapeutically effective amount of Citicoline in association with Folic acid along with the pharmaceutically acceptable excipients useful for the treatment of neurodegenerative disorders.
The solid dosage form is preferably a tablet form prepared by wet granulation process wherein both Citicoline and Folic acid are mixed with appropriate diluents, disintegrants etc and granulated with suitable binding agent, dried, lubricated and finally compressed into tablets.
In another aspect, the present invention describes process for manufacturing Citicoline and Folic acid solid dosage form as follows:
1) Weighing and sifting Citicoline sodium, Microcrystalline Cellulose, Maize Starch and part of croscarmellose sodium through 40# sieve and mixing for sufficient time to get uniform blend;
2) Preparing starch paste using Maize starch and Purified Water;
3) Granulating the blend of step 1 with Starch paste of step 2 to obtain wet mass of suitable consistency;
4) Passing the wet mass through 16 # sieve to obtain wet granules;
5) Drying the granules at 55 C for sufficient period of time;
6) Sifting and sizing dried granules through 18 # sieve;
7) Sifting Folic Acid, Colloidal Silicon dioxide, croscarmellose sodium and Maize starch through 40 # sieve;

8) Mixing dried granules of step 6 and lubricants of step 7 for sufficient period of time.
9) Sifting Magnesium stearate through 40 # sieve and mixing for sufficient time with blend of step 8 to get uniform blend;
10) Compressing the lubricated granules blend on compression machine; and
11) Optionally coating the compressed tablets using film coating solution which contains Hydroxypropyl methyl cellulose (Methocel E5 LV), Diethyl phthalate, Polyethylene Glycol 4000, Talc, Titanium Dioxide, Methylene chloride, Isopropyl Alcohol and colour.
Detailed description of the invention:
The invention will now be described in detail in connection with certain preferred and optional embodiments, so that various aspects thereof may be more fully understood and appreciated.
The present invention describes synergistic pharmaceutical composition in a solid dosage form, preferably a tablet form comprising Citicoline and Folic acid.
The most striking feature of this combination is the requisite specificity of Citicoline and Folic acid as neuroprotective agents. Citicoline achieves therapeutic effect by its ability to increase the synthesis of phosphatidylcholine, the primary neuronal membrane component. Besides it enhances acetylcholine synthesis which ameliorate symptoms caused by loss of cholinergic nerves, Citicoline also influences the outcome of neurodegenerative diseases by its ability to reduce free fatty acid accumulation at the site of injury which prevents further damage. Folic acid improves memory and cognitive functions.
Thus, Citicoline and Folic acid may be administered for long term in elderly patients without any serious side effects. Therefore, Citicoline and Folic acid may be termed as an ideal combination for the management of neurodegenerative or ischaemic cerebral diseases, conditions in which patients suffer till death.

Accordingly, the recommended dose of Citicoline and Folic acid tablet is I tablet twice a day.
According to preferred embodiment, Citicoline is present in an amount of about 200mg to l000mg and Folic acid is present in an amount of 5mg.
In one of the embodiment, the present invention discloses the synergistic pharmaceutical composition comprising Citicoline and Folic acid along with the suitable pharmaceutically acceptable excipients such as diluents, binders, disintegrants, lubricants and glidants; wherein the said pharmaceutical composition is in the form tablet and optionally a film coated tablet.
In another embodiment, the present invention discloses the process for manufacturing fixed dose synergistic pharmaceutical composition comprising Citicoline and Folic acid is prepared as follows:
1. Weighing and sifting Citicoline sodium, Microcrystalline Cellulose, Maize Starch and part of croscarmellose sodium through 40# sieve and mixing for sufficient time to get uniform blend;
2. Preparing starch paste using Maize starch and Purified Water;
3. Granulating the blend of step 1 with Starch paste of step 2 to obtain wet mass of suitable consistency;
4. Passing the wet mass through 16 # sieve to obtain wet granules;
5. Drying the granules at 55 C for sufficient time;
6. Sifting and sizing dried granules through 18 # sieve;
7. Sifting Folic Acid, Colloidal Silicon dioxide, croscarmellose sodium and Maize starch through 40 # sieve;
8. Mixing dried granules of step 6 and lubricants of step 7 for sufficient time;
9. Sifting Magnesium stearate through 40 # sieve and mixing for sufficient time with blend of step 8 to get uniform blend;
10. Compressing the lubricated granules blend on compression machine; and
11. optionally Coating the compressed tablets using film coating solution which contains Hydroxypropyl methyl cellulose (Methocel E5 LV), Diethyl phthalate,

Polyethylene Glycol 4000, Talc, Titanium Dioxide, Methylene chloride, Isopropyl Alcohol and colour.
Diluents are selected from the group consisting microcrystalline cellulose, Maize starch, Lactose, Pregelatinised starch; dibasic calcium phosphate alone or in combination can be used in optimum concentrations for desired product properties.
Binders are selected from purified water, Isopropyl alcohol, alone or in combination along with maize starch, Ethyl cellulose, hydroxy propyl methyl cellulose, povidone K-30 . or like.
Disintegrants such as microcrystalline cellulose, croscarmellose sodium, maize starch, sodium starch glycollate or crospovidone are used alone or in combination.
Lubricants like magnesium stearate, stearic acid and hydrogenated castor oil are used in appropriate concentration, alone or in combination
Glidants such as talc, colloidal silicon dioxide are used alone or in combination.
According to the present invention, the tablet is optionally film coated using (Methocel E5 LV), hydroxyl propyl methyl cellulose along with plasticizer, Diethyl phthalate, Polyethylene Glycol 4000, talc, titanium dioxide Methylene chloride, Isopropyl Alcohol and colour.
The invention can be illustrated by following non limiting examples;
Example 1:
CITICOLINE AND FOLIC ACID TABLETS (lOOOmg + 5mg)
Label Claim : Each film-coated tablet contains:
Citicoline Sodium equivalent to Citicoline 1 000mg
Folic Acid IP 5mg
Excipients q.s.

Sr.No Ingredients Qty/ Tablet Range
1. Citicoline Sodium equivalent to Citicoline 1000.00 mg
2. Folic Acid 5.00 mg
3. Microcrystalline Cellulose 38.59 mg 25 mg- 55 mg
4. Maize Starch 90.00 mg 75 mg- 105 mg
5. Croscarmellose Sodium 45.00 mg 35 mg-55mg
6. Colloidal Silicon Dioxide 10.00 mg 7mg- 13 mg
7. Magnesium Stearate 18.00 mg 15 mg- 20 mg
Example 2:
CITICOLINE AND FOLIC ACID TABLETS (750mg + 5mg)
Label Claim : Each film-coated tablet contains:
Citicoline Sodium equivalent to Citicoline 750mg
Folic Acid IP 5mg
Excipients q.s.

Sr
Nc > Ingredients Qty/ Tablet Range
1. Citicoline Sodium equivalent to Citicoline 750.00 mg
2. Folic Acid 5.00 mg
3. Microcrystalline Cellulose 13.10 mg 10 mg -20mg
4. Maize Starch 10.00 mg 5mg- 15 mg
5. Croscarmellose Sodium 32.00 mg 25 mg -35 mg
6. Colloidal Silicon Dioxide 8.00 mg 6mg- 10 mg
7. Magnesium Stearate 14.00 mg 10 mg- 16 mg

Example 3:
CITICOLINE AND FOLIC ACID TABLETS (500mg + 5mg)
Label Claim : Each film-coated tablet contains:
Citicoline Sodium equivalent to Citicoline 500mg
Folic Acid IP 5mg
Excipients q.s.

SrNo Ingredients Qty/Tablet Range
1. Citicoline Sodium equivalent to Citicoline 500 mg
2. Folic Acid 5mg
3. Microcrystalline Cellulose 200 mg 180mg-220mg
4. Maize Starch 90 mg 75 mg- 105 mg
5. Croscarmellose Sodium 30 mg 25 mg-35 mg
6. Colloidal Silicon Dioxide 8mg 6 mg- 10 mg
7. Magnesium Stearate 14 mg 10 mg- 16 mg
Example 4:
CITICOLINE AND FOLIC ACID TABLETS (200mg + 5mg)
Label Claim : Each film-coated tablet contains:
Citicoline Sodium equivalent to Citicoline 200mg
Folic Acid IP 5mg
Excipients q.s.

Sr.
No Ingredients Qty/Tablet Range
1. Citicoline Sodium equivalent to Citicoline 200.00 mg
2. Folic Acid 5.00 mg
3. Microcrystalline Cellulose 78.00 mg 60mg-100mg

4.
Maize Starch 35.89 mg 20mg-50mg
5. Croscarmellose Sodium 12.80 mg 7mg- I7mg
6. Colloidal Silicon Dioxide 3.20 mg 2 mg - 5 mg
7. Magnesium Stearate 5.60 mg 2mg-7mg
The stability of the said composition is carried out according to ICH guidelines. The product of the said composition is stable for 6 months at accelerated conditions of 40°C -75% RH and at real time conditions of 30°C-65 % RH.

We Claim,
(1) A novel pharmaceutical solid oral dosage form comprising Citicoline in an amount of 200mg tolOOOmg and Folic acid in an amount of 5mg along with pharmaceutically acceptable excipients such as diluents, binders, disintegrants, lubricants, glidants; useful for the treatment of neurodegenerative disorders.
(2) The novel pharmaceutical solid oral dosage form as claimed in claim 1, wherein the diluents used are microcrystalline cellulose, Maize starch, Lactose, Pregelatinised starch; dibasic calcium phosphate either alone or in combination.
(3) The novel pharmaceutical solid oral dosage form as claimed in claim I, wherein the binders are selected from purified water, Isopropyl alcohol, alone or in combination with maize starch, Ethyl cellulose, hydroxy propyl methyl cellulose, povidone K-30 or like.
(4) The novel pharmaceutical solid oral dosage form as claimed in claim 1, wherein the disintegrant used are microcrystalline cellulose, croscarmeUose sodium, maize starch, sodium starch glycollate or crospovidone either alone or in combination.
(5) The novel pharmaceutical solid oral dosage form as claimed in claim 1, wherein the lubricants used are magnesium stearate, stearic acid and hydrogenated castor oil are used in appropriate concentration, alone or in combination.
(6) The novel pharmaceutical solid oral dosage form as claimed in claim 1. wherein the glidants used are talc, colloidal silicon dioxide alone or in combination.
(7) The novel pharmaceutical solid oral dosage form as claimed in claim 1, wherein the composition is optionally film coated using (Methocel E5 LV), hydroxyl propyl methyl cellulose along with plasticizer-, Diethyl phthalate, Polyethylene Glycol 4000, talc, titanium dioxide Methylene chloride, Isopropyl Alcohol and color.
(8) The novel pharmaceutical solid oral dosage form as claimed in claim 1, wherein process for preparing said composition comprises steps of:
a. Weighing and sifting Citicoline sodium, Microcrystalline Cellulose,
Maize Starch and part of croscarmeUose sodium through 40# sieve and
mixing for sufficient time to get uniform blend;
b. Preparing starch paste using Maize starch and Purified Water;

c. Granulating the blend of step (a) with Starch paste of step (b) to obtain wet
mass of suitable consistency;
d. Passing the wet mass through 16 # sieve to obtain wet granules;
e. Drying the granules at 55 C for sufficient period of time;
f. Sifting and sizing dried granules through 18 # sieve;
g. Sifting Folic Acid, Colloidal Silicon dioxide, croscarmellose sodium and
Maize starch through 40 # sieve;
h. Mixing dried granules of step (f) and lubricants of step (g) for sufficient
period of time;
i. Sifting Magnesium stearate through 40 # sieve and mixing for sufficient
time with blend of step (h) to get uniform blend;
j. Compressing the lubricated granules blend on compression machine; and k. Optionally coating the compressed tablets using film coating solution
which contains Hydroxypropyl methyl cellulose (Methocel E5 LV),
Diethyl phthalate, Polyethylene Glycol 4000, Talc, Titanium Dioxide,
Methylene chloride, Isopropyl Alcohol and colour.