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This invention relates to a formulation for iron-chelation The formulation of the present invention is useful for treating patients suffering from the disease of Thalassemia .
This invention relates particularly to a formulation useful fat the treatment of patients suffereing from the disease of Thalassemia with increased therapeutic efficacy.
Thalassemia is a dreaded disease among children because of genetic disorder. The disease is caused due to hereditary disorders connected with defective haemoglobin synthesis, characterised by hypochromia, microcytosis, haemolysis and a variable degree of anaemia. Thalassemia involves a heterogeneous group of molecular defects and present with a wide spectrum of clinical expressions.
Patients sufferening form Thalassemia suffer from anaemias with decreased or absence of synthesis of a globin chain of a normal haemoglobin. The patients of thalassemia are broadly classified into two major groups according to the affected globin chain. The patients suffereing form Alpha (
Rarely one may find also patients suffereing form Thalassemia delta (£) and Gamma (V) chain disorders, as well as those associated abnormal hamoglobin structure (e.g. Hb Lepose and Hb Constant Spring) . Such types of disease which also contribute to the Thalassemia syndromes.
The disease Thalassemia occurs world wide with a particular high incidence in the Mediterranean basin and in the South-East
Asia. Malaria is also endemic in these areas - a significant fact since indirect evidence suggests that- Thaiassemia (major) heter-ozygosity confers protection against malaria.
The P -Thaiassemia or Thaiassemia major type of the disease
comprises a heterogeneous group of disorder usually characterised
0 +
by absence of (P ) or decreased (P ) globin synthesis.
The type of P Thaiassemia is also classified according to the severity of the anaemia. These clinical classification serves to differentiate homozygous (Thaiassemia intermedia or Thaiassemia. major) from heterozygous state {Thaiassemia minima or Thaiassemia minor). Though it does not reflect genetic mutations, Thaiassemia (minor) is a reduced rate of P -globin synthesis, with an increased d-p globin chains, but it not threat to a normal life
In case of Thaiassemia (major) also known as Cooley's anaemia, Mediterranean anaemia and Von - Jacksch*s anaemia is characterised by marked anaemia (ranging from 1 to 6 gm/dl of hamoglobin), severe hemolysis and ineffective erthropoiresis. The diagnosis is made in the 1st year of the life of the patient, often as early as 3 months old baby. In the case of Thaiassemia (major), iron in the haemoglobin also breaks down and gets deposited in the vital organs of the body of the patients e.g. liver, kidney, spleen etc. This is also known as iron overloading in the body and the life span of the child suffering from Thaiassemia (major) becomes unpredictable. Every year out of i,00,OOO children born with Thaiassemia (major) in the world, 10,000 are born in India.
The method which is available hitherto for the treatment of
Thalassemia (major) is by the life long blood transfusion coupled with the taking of the drug called 'Desferal' daily intermuscula-ry. The chemical name of the drug is Deferoxamine Methane Sulpho-
nate and the chemical formula is C H NO S.
28 52 4 11 Presently followed therapy is giving injection of Desferal for
the excretion of iron from the body of the patient through urinay excretion. It was found by medical profession that Desferal had many side effects like swelling of limbs, stiffness in joints and may inhibit a. number of tumour eel 1 prol iferation .parasite growth and the proliferation of the cerebral Malarial Parasite, Plasmod-
_--£ ^^
ium (Fal i parura.)
In addition , the above drug is to be injected daily under the skin of the patient in a controlled manner in such a way to avoid any side reaction causing allergic conditions .Such a treatment is M highly painful . The treatment is also costly as the vial containing 500 mg of dry active substance will cost Rs ISO/ each. Though a new oral iron chelating drug known as L-1,CP20,DMPH, Deferiprone (Chemical Name,1,2 Dimethyl 3 Hydroxyp-yridine 4-one ) has been reported very recently .It is yet to establish its potentiality of patients suffereing from the disease of Thalassemia . The reported side effects of this drug are 1. Myelotoxicity ie occurance of neutropenia 2. Orthropathy ie skeleto muscular pain and swelling around knee and hipjoints and lastly mild zinc defficiency occasionally leading to dermatopathy (Reference may be made to the Proceedings of National Thalassemia Conference, 5-6 February, 1994, held in Delhi; LI: Oral Iron Chelation Therapy- Indian Study by M .B. Agrawal). The cost of a capsule Deferiprone is approximately Rs
12/-. A patient has to take 3-6 capsules per day. This will amount to Rs 10OO- 2000/- per month.
Another method which is available for the treatment of the disease Thalassemia (major) is by the bone marrow transplantation (BUT). This is done by the taking the bone marrow of the matching donor and injecting it to the patient. The cost of such a treatment is around Rs 15,00,000/-. This is therefore beyond the reach of common man. Several treatments using bone marrow have been performed in Italy. Recently Christian Medical College (Vellore) and Appolo Hospital (Madras) have also started this type of BMT treatment in India, but the cost is also on the higher side (Rs 7 Lac approx). Further increase in life span expectancy of these patients have yet to be established.
Under the prevailing present day conditions, regular blood transfusion and use of the drug 'Desferal1 by injection as explained above, is the best way of treatment of this dreaded disease. Since a. decade, efforts are being made by medical profession throughout the world to find a treatment of this disease by a drug which will be low cost and can be administered orally and have no side effect, but there has been no success so far.
The main objective of the present invention is to provide a formulation useful for iron-chelation for the treatment of Thalassemia.
Another objective of the present invention is to provide a formulation useful for the treatment of Thalssemia with increase therapeutic efficacy.
Yet another of the present invention' objective is to provide
a formulation for the treatment of Thaiassemia which is much cheaper and hence afforadable by a common man. .
Still another objective of the present invention is to provide a formulation useful for the treatment of patients suffering from Thalassemia which can be administered orally.
Still yet another object of the present invention is to provide a formulation whic has no side effect and is very convenient to administer.
Another object of the present invention is to provide a formulation useful for the treatment of Thalassemia, the dose of which can be continued for a long period without damaging any vital organ of the body of the patients.
With the above objectives in view our work initially directed in red blood cells and at the same time, to enhance the oxygen carrying capacity to the tissues. Different compounds of Vanadium, Arsenic, oxalic acid and citric acid were tried initially but these compound: fai1ed in making a major break through. BY these research only sodium meta vanadate and arsenic were found to be effective partially in increasing the red blood cells and also helped in increasing the percentage of haemoglobin in the blood of the patients suffering from Thalassemia.
It has been observed that patients suffering from Thalassemic disease have a change in complexion, anorexia, blackness of gums, increase in serum ferritin level and a remarkable iron overload in the body due to the breaking down of red blood cells. The treatment mentioned above which involves repeated blood transfusion also accumulates the above elements in the body of the patients suffering from Thalassemic disease.
Children suffering from Thai lassemia diesease dolnoVshow any
resistance from viral infection and also suffer from body ache. / They are also immune to malaria.
Our reserach work was primarilly directed to find an immediate solution for the survival of suoh patients, whfl hftd An ir-fm
overload in the body and the other side ailments mentioned above. Efforts, therefore, were directed towards finding out an alternate drug which can be used orally and have no side effects. Anemonin Pretensis (powder) has been used for many years by tribals of Siberia to poison their arrows. Anemonin Pretensis is extracted with an organic solvent such as ethanol from fresh whole wind flower plant with roots and flowers and some fruits of wind flowers. The dried material is treated with an organic solvent such as ethanol and refluxed. Anemonin Pretensis obtained from the extraction is filtered and recrystalIized from an organic solvent such as ethanol.The medical history of this herb also reveals that this herb was involved in homeopathic practice in 1805 for some female hormone problem.
th'emonin Pretensis) is a pure herbal product . This is
present in the wind flower plant which is growing in wild state in the open fields and plains in many parts of Europe, Russia and Turkey in Asia. The chemical formula of Anemonin prentesis having the formula 1, shown in the drawing accompanying this
specification, isolated from fresh wind flower plant is 1-2
dihydroxy 1-2 cyolobutane diacrylic acid dilactone. Reference may
th be made to Merk index p-87, Entry no. 677, 9 edition .
Patients suffering from Thalassemic disease are immune to
malarial attack. Considering the nature and considering that quinine sulphate a known antimalarial drug which has antipyretic and analgesic properties, we considered incorporating quinine sulphate in the formulation to reduce the high temperature and body-ache problem of the patients suffering from Thalassemic disease.
Quinine is having the chemical composition C H N 0 . It is
40 48 4 4 obtained from Cinchona bark available in India, Srilanka, Equa-
dor, Columbia, Peru and Bolivia. Cinchona Thrives at higher elevation such as 6000-6500 ft. Quinine is also extracted with an organic solvent such as ethanol from the bark of the plant (Cinchona), refluxed, filtered and recrystal1ized with an organic solvent such as ethanol. Quinine sulphate is prepared by reacting quinine and dilute sulphuric acid in a molar ratio. It is a snow white, light odourless extremely bitter crystallized needles
having the chemical composition C H N 0 , H SO , & H 0. Refer
40 48 4 4 24 2
th
ence may be made to Merk Index p-1049, Entry no. 7879, 9 edition.
By our continuous and sustained reserach work based on the above mentioned directions we observed that when we blended the powder of Anemonin Pretensis with quinine sulphate and dissolved in suitable solvent and the solution was administered orally td the patient suffering from Thalassemia, there was remarkable improvement of complexion and reduction of serum ferritin level. There was no fever . There was also no body ache . The treatment was continued and more patients were put on trial on this drug. There were no side effects. This revealed that the fomulation is very useful in te treatment of the patients suffering fora
Thalassemia.
Accordingly the present invention provides a pharmaceutical formulation useful as low cast iron chelating drug for Thalassemia patients which comprises:
i) 0.02 to 0.12 wt% powder of Anemonin Pretensis , ii) 0.0005 to 0.003 wt% quinine sulphate , iii) \0 to 40wt% distilled or demineralised water iv) 100 to 60 wt% edible solvent.
The solvent used may be selected from solvent ethanol, absolute alcohol etc.
The Anemonin pretensis powder and the quinine sulphate employed in the formulation may be of pharmaceutical grade.
The Anemonin pretensis possesses the properties of chelating iron. Quinine sulphate seems to accelearate the chelation of iron present in the body of the patient suffering from thalassemia. We have found by forming a formulation of Anemonin pretensis and quinine sulphate in the amount mentioned above results in an unexpected properties which ar not present in the individual components. There is therefore a synergistic activity when they are combines in the above mentioned quantities.
The formulation of the present invention is therefore not a mere admixture of the ingredients employed but a synergistic mixture, the properties of which are not merely the aggregate properties of the individual ingredients.
The formulation of the present invention when administered to patients suffering from Thalassemic dbisease works as follows: The iron present in the body of the patients forms a complex with anemonin shown in the formula 2 (Closed ring structure) or in formula 3 (Streched structure) where three molecules of anemonin can form complex with two iron atoms. The quinine sulphate present in the formulation acts as a catalyst for the formation of the complex thereby not only reducing the amount of Anemonin in the composition and but also use the maximum amount of iron present in the body. In addition the antipyretic and analgesic property of quinine sulphate helps to control the fever and the bodyache problem present in such patients.
The iron complex so formed is soluble in the body fluids. The fluid containing the complex reaches the kidney through the body system of the body and it is then excreted from the body of patients through urine. Some of the complex is also excreted through elementry canal and finally facaeces. By this processor the excess iron present in the body of the patients is removed thereby enhancing the life span of the patients.
The following examples are given by way of illustration and should not be construed to limit the scope of the present invention: Example 1.
A formulation was prepared by blending the following ingredients: Powder of Anemonin Pretensis O.O2 wt % of the formulation
Quinine Sulphate O.OO05 wt X of the formulation
Demineralised Water 10 wt * of the formulation
Ethanol Solvent 9O wt X of the formulation
Example 2
A formulation was prepared by blending the following ingre-d ients:
Powder of Anemonin Pretensis O.O4 wt % of the formulation
Quinine Sulphate O.OO08 wt % of the formulation
Demineralised Water 10 wt % of the formulation
Ethanol Solvent 90 wt X of the formulation
Example 3
A formulation was prepared by blending the following
ingredients:
Powder of Anemonin Pretensis O.O6 wt % of the formulation
Quinine Sulphate O.O01 wt % of the formulation
Demineralised Water 1O wt X of the formulation
ethanol Solvent 90 wt % of the formulation
Example £
A formulation was prepared by blending the following ingredients :
Powder of Anemonin Pretensis 0.08 wt % of the formulation
Quinine Sulphate 0.002 wt % of the formulation
Demineralised Water 10 wt % of the formulation
ethanol solvent 90 wt % of the formulation
Exampl e 5^
A formulation was prepared by blending the following
ingredients:
Powder of Anemonin Pretensis O.O9 wt % of the formulation
Quinine Sulphate Demineralised Water Ethanol Solvent Example 6
O.OO1 wt % of the formulation 10 wt X of the formulation 90 wt X of the formulation .
A formulation was prepared by blending the followimg ingredients.
Powder of Anemonin Pretensis 0.12 wt X of the formulation
Quinine Sulphate O.OO3 wt X of the formulation
Demineralised Water 10 wt % of the formulation
Ethanol solvent 90 wt X of the formulation
Example 7_
A formulation was prepared by blending the following ingred ients:
Powder of Anemonin Pretensis 0.09 wt X of the formulation
Qunine Sulphate O.O01 wt X of the formulation
Demineralised Water 80 wt X of the formulation
Ethanol solvent 20 wt X of the formulation
Exampl e 8_
A formulation was prepared by blending the following ingredients :
Powder of Anemonin Pretensis O.08 wt X of the formulation
Qunine Sulphate O.O01 wt X of the formulation
Demineralised Water 40 wt X of the formulation
Ethanol Solvent 6O wt X of the formulation
The formulations of the present invention prepared according the above mentioned examples were administered to a number of
patients suffering from Thalassemia. Iron chelation upto 9OX can be achieved by the formulation of the present invention which en be confirmed from the results shown in Table I.
Table - I Efficacy of the formulation of the present invention.
(Table Removed)
The main advantages of the formulation of the present invention are:
1. Iron-chelation using the formulation of Anemonin
Pretensis and Quinine sulphate is upto 90 X.
2. The cost of the formulation is around Rs 150-20O/- per 30 ml vial
and it can be used for one month. The cost is much lower as
compared to other currently available medicines in the market for
the treatment of Thalassemia.
3. The formulation has no toxic' effects even when it is
administered for longer period.
4. The formulation is tasteless and odourless and can be administered orally.
We Claim:
1. A pharmaceutical formulation useful as low cast iron chelating drug for
Thalassemia patients which comprises:
i) 0.02 to 0.12 wt% powder of Anemonin pretensis , ii) 0.0005 to 0.003 wt% quinine sulphate , iii) 40 to 40wt% distilled or domaterialized water |Mo iv) 100to60wt% edible solvent.
2. A pharmaceutical formulation as claimed in claims 1 wherein the
Anemonin pretensis powder used is of pharmaceutical grade.
3. A formulation as claimed in claims 1-2, wherein the solvent used is
selected from ethanol or absolute alcohol.
4. A formulation as claimed in claims 1-3 wherein quinine sulphate used is
of pharmaceutical grade.
5. A process for the preparation of a pharmaceutical composition as claimed
in claims 1 -4, which comprises mixing by conventional method
i) 0.02 to 0.12 wt% powder of Anemonin pretensis, ii) 0.0005 to 0.003 wt% quinine sulphate, iii) 40 to 40% distilled or demineralised water frio iv) 100 to 60 wt% edible solvent.
6. A process for the preparation of a pharmaceutical composition low cast
iron chelating drug for Thalassemia patients substantially as herein
described with reference to the examples.
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