WO2011028098A1

WO2011028098A1 - Product and method for treating thrombocytopenia

Info

Publication number: WO2011028098A1
Application number: PCT/NL2009/050529
Authority: WO
Inventors: Abul Bashr Mohammed Fazlul Karim; Choudhury Mahmood Hasan; Hossain Sohrab; Muhammed Abdullah Al-Mansur; Refaat Bari Karim
Original assignee: Abul Bashr Mohammed Fazlul Karim; Choudhury Mahmood Hasan; Hossain Sohrab; Muhammed Abdullah Al-Mansur; Refaat Bari Karim
Priority date: 2009-09-03
Filing date: 2009-09-03
Publication date: 2011-03-10

Abstract

Object of the invention is providing a product and method for increasing the number of platelets in blood. This object is met by providing a pharmaceutical composition for treating thrombocytopenia, which pharmaceutical composition comprises a pharmaceutically acceptable carrier and an active compound composition comprising one or more compounds present in papaya leaf dry matter chosen from the group consisting of papain, chymopapain, alkaloids, benzylglucosinolate and tannins.

Description

Title: Product and method for treating Thrombocytopenia

The invention is directed to a product and method for treating thrombocytopenia.

Platelets are small cytoplasmic bodies derived from cells. Platelets are also called thrombocytes. They circulate in the blood of mammals and are involved in hemostasis leading to the formation of blood clots. Platelets are produced in blood cell formation (thrombopoiesis) in bone marrow, by budding off from megakaryocytes. The platelet count in the circulating blood of a healthy person is typically between 150 · 10⁹ and 400 · 10⁹ per liter of blood. Newborn babies have a slightly lower level, but are normally within the adult range by three months of age.

Thrombocytopenia is a condition in which there is a deficient number of circulating platelets in the blood. The term "blood" as used herein refers to peripheral blood, i.e. the blood circulating throughout the body. The platelet count in the blood of a person with thrombocytopenia is typically reduced to number below the normal lower limit of 150 · 10⁹ platelets per liter blood. Moderate thrombocytopenia may be defined as a platelet count between 60 · 10⁹ and 100 · 10⁹ per liter blood. Severe thrombocytopenia may be defined as a platelet count below 60 · 10⁹ per liter blood.

The main effect of a reduced platelet count is an increased risk of bleeding. If a person's platelet count is below 50 · 10⁹ per liter blood, bruising with minor trauma may be expected; if it is below 30 · 10⁹ per liter blood, spontaneous bruising will be seen There is a particularly high risk of spontaneous bleeding once the platelet count drops below 15-30 · 10⁹ per ml. Furthermore, thrombocytopenia may increase the risk of hemorrhage, for instance from the nose or at the retina. The most serious complication, which is potentially fatal, is spontaneous bleeding inside the head (intracranial) or in the gut (gastrointestinal). A person with Thrombocytopenia may also complain of malaise, fatigue and general weakness, with or without accompanying blood loss. Thrombocytopenia may be caused by defective platelet production in the bone marrow or by increased destruction of the platelets. In addition, thrombocytopenia may also be medication induced. For example, many chemotherapeutic agents are cytotoxic and depress megakaryocyte production. This type of thrombocytopenia is called chemotherapy induced thrombocytopenia (CIT). Ingestion of large quantities of alcohol has a similar marrow-depressing effect leading to transient (temporary) thrombocytopenia. Since recent chemotherapy drugs are not only more potent and effective, but also highly toxic, thrombocytopenia has recently been encountered more frequently in patients treated with chemotherapeutic agents.

Thrombocytopenia is usually treated by eliminating the underlying problem, e.g. by discontinuing suspected drugs that cause thrombocytopenia or treating underlying sepsis. However, discontinuing drugs may be very undesirable when treating patients for life-threatening diseases such as chemotherapy. In these cases, fresh blood or platelet rich transfusion is often used to increase the platelet count in a patient. Disadvantages of using transfusions are many, for example the risk of fever, viral infections and blood contamination, the need of a suitable blood donor and high medical costs. Furthermore, it is believed by many clinicians that the transfused platelets survive no longer than the patients' own platelets. Transfused platelets are thus rapidly destroyed in the blood of the recipient and the benefit of transfusion may therefore not be very significant.

Regarding medicinal treatment of thrombocytopenia, not many suitable drugs are known. However, a drug called Neumega (Oprelvekin) is approved recently in the United States for prevention of chemotherapy induced thrombocytopenia. Disadvantage of this drug is that it has to be administered by injection, should be kept at low temperatures and has a number of side effects, such as fever and irregular heartbeats. In addition, Neumega is also very costly and not available in many developing countries. It is an object of the present invention to solve at least in part the problems encountered in the prior art regarding the treatment of thrombocytopenia.

In a first aspect, this object is met by providing a pharmaceutical composition for treating thrombocytopenia, which pharmaceutical composition comprises a pharmaceutically acceptable carrier and an active compound composition comprising one or more compounds present in papaya leaf dry matter chosen from the group consisting of papain, chymopapain, alkaloids, benzylglucosinolate, tannins , flavonoids and perhaps many others.

The inventors surprisingly found that a mammal suffering from thrombocytopenia, in particular a mammal suffering from CIT, can successfully be treated by administration of an active compound composition according to the invention, in particular by administration of papaya leaf dry matter. The advantage of treating mammals suffering from thrombocytopenia by administration of the active compound composition of the invention is that this composition is a relatively harmless product with few side effects that is easy to come by and can easily be obtained from papaya leaves.

When administered to a mammal suffering from thrombocytopenia, in particular CIT, the active compound according to the invention has a positive effect on the platelet count in the blood of the mammal. It is believed, without wishing to be bound by any theory, that this active compound composition stimulates and/or improves the megakaryocytes to produce sufficient numbers of platelets to maintain a suitable platelet count in mammals, in particular during chemotherapy.

The pharmaceutical composition may be used for treating any mammal, but was in particularly found suitable for treating humans.

The active compound composition according to the invention can be found in papaya leaf dry matter. As used herein, the term "papaya leaf dry matter" refers to all compounds found in papaya leaves excluding any water molecules. Papaya leaf dry matter may be obtained by drying papaya leaves, for example by freeze drying papaya leaves. A papaya leaf as used herein is defined as a leaf from the plant Carica papaya.

The active compound composition according to the invention may comprise one or more compounds present in papaya leaf dry matter. The group may consist of vitamins, biologically active compounds, alkaloids and any other biologically active chemical compounds.

Vitamins present in papaya leaf dry matter are β-carotene, thiamin (vitamin Bl), riboflavin (vitamin B2), niacin (vitamin B3), vitamin B6, ascorbic acid (vitamin C).

Examples of metals present in papaya leaf dry matter are calcium, iron, magnesium, potassium and sodium.

Examples of biologically active compounds present in papaya leaf dry matter are proteolytic enzymes, for example papain and chymopapain. Papain and chymopapain aid in the digestion of protein. These two compounds may contribute to the positive effects on the platelet count, e.g. by a protein digestion process. Therefore, the active compound composition may comprise papain, chycopapain or both.

Examples of alkaloids present in papaya leaf dry matter are carpaine, nicotine, myosmine, piperidine, dehydrocarpaines, pseudocarpaine. Alkaloids often have pharmacological effects and are used as medications and recreational pharmaceutical compositions. One or more of the alkaloids may therefore play an important role in the thrombocytopenia treatment according to the method of the invention. Thus, the active compound composition may comprise one or more alkaloids, more preferably one or more compounds chosen from the group consisting of carpaine, nicotine, myosmine, piperidine, dehydrocarpaines and pseudocarpaine.

Examples of other chemical compounds present in papaya leaf dry matter are flavonols (such as kaempferol, quercetin, myricetin and fisetin), flavonoids, benzyl glucosinolate, tannins, vitamin A equivalents and phosphorus. Flavonaols or flavonoids are known to be used as ingredients in many pharmaceuticals compositions. The active compound may thus comprise flavonaols, flavonoids or both.

Glucosinolates are well known for their toxic effects in both humans and animals at high doses. However, at subtoxic doses, they may act as chemoprotective agents against chemically-induced activities. The active compound composition may comprise glucosinolates, in particular benzyl glucosinolate..

Tannins are astringent, bitter plant polyphenols that either bind and precipitate or shrink proteins. Tannins are large polyphenolic compounds containing sufficient hydroxyls and other suitable groups (such as carboxyls) to form strong complexes with proteins and other macromolecules. The complex- forming properties of tannins may contribute to the positive effects on the platelet count in blood after administration of the active compound composition. Thus, the active compound composition may comprise tannins.

The active compound composition may be more accurately identified, e.g. by testing the effect of the different compounds described hereinabove on thrombocytopenia.

It would be desirable to isolate the active compound composition from the papaya leaves. This could help when further optimizing the pharmaceutical composition. In case the active compound composition would comprise more than one active compound, the ratio of these compounds could be varied to further optimize the pharmaceutical composition.

The pharmaceutical composition according to the invention may be used for the treatment of thrombocytopenia, in particular for the treatment of medicine induced thrombocytopenia, more in particular for the treatment of CIT. The pharmaceutical composition according to the invention works particularly well for treating chemotherapy induced thrombocytopenia. It can be administered in combination with chemotherapy drugs and has shown very good results in patients of different ages and gender (see also the examples hereinbelow). The pharmaceutically acceptable carrier may refer to a diluent, adjuvant, excipient or carrier with which a compound of the invention is administered. Examples of suitable carriers are well known in the art and may include, but are not limited to, any of the standard pharmaceutical carriers.

The pharmaceutical composition described hereinabove may be used in a method for treating a mammal having a deficiency in platelets in the blood by administration of said pharmaceutical composition.

Administration of the active compound composition may be conducted in various ways, for example by injection or oral intake. Preferably, the active compound composition is administered by oral intake, because of the relative ease and comfort for the patient. Preferably, the pharmaceutical composition in the method according to the invention is papaya leaf dry matter. In this case, a daily dosage of at least 50 mg, preferably at least 100 mg, e.g. a dosage of 132 mg of papaya leaf dry matter is administered to patients with moderate thrombocytopenia. In cases of severe thrombocytopenia, these dosages may be 1.5 times higher. Because papaya leaf dry matter is relatively harmless and has few side effects, the maximum daily dosage may be as high as 1000 mg or even higher. It is desirable to divide the daily dosage in smaller dosages, preferably two or three, that are administered over the course of a day. For example, a treatment according to the method of the present invention may be administering to a patient for several days a total daily dosage of 132 mg by giving the patient a first dosage of 66 mg, in the afternoon followed by a second dosage of 66 mg in the evening.

When determining the daily dosage needed for a patient, one should take into account the body weight of the patient, as well as the gender and age of the patient.

The pharmaceutical composition according to the invention may further be used as a prophylactic. Such a prophylactic may for example be administered to a patient receiving chemotherapy to decrease the chances of the patient of developing CIT. The pharmaceutical composition of the invention may be prepared according to a method comprising the steps of:

- providing papaya leaves, preferably fresh matured papaya leaves; and

- drying said papaya leaves, preferably by freeze drying, thus obtaining papaya leaf dry matter.

Preferably, freshly matured papaya leaves are used for the preparation of the pharmaceutical composition, because such leaves typically contain the highest amounts of papaya leaf juice. It is expected, without wishing to be bound by any theory, that the highest amounts of the active compound composition can be found in papaya leaf juice. The term "papaya leaf juice" as used herein is defined as any liquid present in a papaya leaf, including any compounds that may be dissolved in said liquid.

Drying of the papaya leaves may be done in various ways. Preferably however, the papaya leaves are freeze dried.

Prior to drying, the papaya leaves may be washed and/or disinfected, e.g. from bacteria. Papaya leaves may for example be washed with water. Papaya leaves may also be soaked in a sodium benzoate solution, which may function as a bacteriostatic and/or fungistatic. This is preferably done at a temperature of 75-100 °C, more preferably 85-98 °C. A suitable concentration of sodium benzoate concentration may be 0-2 wt.%, for example about 1 wt.% sodium benzoate. Soaking in such a solution may be conducted for 1-10 minutes, preferably 1-2 minutes.

After drying, the papaya leaf dry matter may be grinded into granules. Grinding can be done in many different ways, such as squeezing, pressing or crushing. Thus obtained granules are easy to handle and facilitate the use of the papaya leaf dry matter in possible further steps.

The obtained papaya leaf dry matter according to the method of the invention may be made into a suitable pharmaceutical composition by applying a pharmaceutically acceptable carrier. This can for example be established by encapsulating papaya leaf dry matter, which may be in the for of granules obtained from a grinding process. A capsule obtained according to the method of the invention and the additional steps of freeze-drying and encapsulation may comprise 10 - 2000 mg of said granules, preferably 80 - 500 mg of said granules, more preferably 150 - 350 mg of said granules.

The present invention will be further illustrated by the following examples.

Example 1: Preparation of the Pharmaceutical Composition

Freshly matured papaya leaves were collected and then washed thoroughly. The washed leaves were then soaked in water containing 1% sodium benzoate for 1 to 1.5 minutes at 90-95 °C. The thus obtained soaked leaves were then freeze dried and grinded into granules. In case the freeze dried leaves were not dry enough, the freeze- dried leaves were blended. The granules were then passed through a sieve of 15-20 mm mesh. The sieved granules were encapsulated into 1 or 2 sized capsule shells. Each capsule contained 250 mg papaya leaf granules.

Example 2: Treatment of Humans suffering from CIT:

Some Case Records are now included to show the effectively of the product in treating Chemotherapy Induced Thrombocytopenia (CIT). Capsules were made according to the method of example 1, comprising 66 mg papaya instead of 250 mg. These capsules are referred to as Platelet Boost (PB) Capsules.

Fifty- six human patients were treated for thrombocytopenia by oral administration of Platelet Boost Capsules. Fifteen of these patientsl5 had moderate and forty-one patients had severe degree of thrombocytopenia. Moderate thrombocytopenia in all 15 patients improved by administration of the Platelet Boost Capsules in 2 divided doses for a total daily dosage of 132 mg. All patients underwent peripheral blood count every 3 days or sometimes every day. Improvement in the platelet count was noticed in 3 to 5 days. 31 patients had severe thrombocytopenia with platelet count between 30,000 and 55,000 per mm³ of peripheral blood. The details of some of these patients are noted below: 1. SMR: aged 34 yrs: Male had metastatic (Liver) Colorectal Cancer and was advised Folfox 4 abroad. He already had about 4 cycles of Folfox 4 and although was planned 4 more cycles, he could not get chemotherapy due to severe thrombocytopenia. He was treated with Platelet transfusion but the benefit was transient. He was given Platelet Boost Capsules as stated above and his platelet count improved in a week. When reassessed, no liver metastasis could be detected. In view of the good response of Folfox 4, it was decided to continue the same chemotherapy. He was doing quite well for three months (3 cycles of Folfox 4 were administered) and no thrombocytopenia was noted any time during the period. Unfortunately when reassessed, floried liver metastases were noted and the patient did not come for any further therapy due to frustration.

2. NN: Female aged 38 yrs. She was undergoing post-operative adjuvant chemotherapy for T2N2M0 poorly differentiated duct cell carcinoma of the breast with Docetaxel, Doxorubicin and 5 fluorouracil in each cycle. She used to get G-CSF, at least 2 injections on alternate day subcutaneously after the chemo-schedules. After the 3rd cycle leucocytes and thrombocytes count went down and she recovered quickly from leucopenia. She got 2 platelet transfusions with transient benefit. When the Platelet Boost Capsules were given for 5 days, her thrombocyte count returned to reasonable level (about 110,000/mm3). She was given for all subsequent chemo cycles, prophylactic Platelet Boost she was not further troubled by thrombocytopenia.

3. MR: 10 years: A female child was referred to us post operatively for Non-Hodgkin's Lymphoma (NHL) of the small intestine which produced obstruction of the gut. During the 3rd cycle of the CHOP she developed myelosuppresive signs with both leucopenia and thrombocytopenia. Leucopenia improved with granulocyte colony- stimulating factor (G-CSF) injections but thrombocytopenia improved from 60,000/mm3 to 240,000/mm3 only after the Platelet Boost Capsules were ingested only for 3 days. She has now completed CHOP therapy with platelet boost capsules and is apparently free from NHL at present.

4. AMH: 58 years male suffering from inadequately operated pancreatic carcinoma and presented with a mass (± 8x8 cm2) in the epigastrim but no jaundice. He improved spectacularly with aggressive chemotherapy including Gemcitabine, Docetaxel, 5-FU & Oxaliplatin and the mass disappeared completely but he developed Leucopenia and thrombocytopenia after 7 cycles of chemotherapy. As usual leucopenia improved but thrombocytopenia persisted with 5 platelet transfusions. It improved gradually and rather slowly when he started taking the Platelet Boost capsules. His platelets returned to the normal level after about 10 days.

5. AIK: 60 years: Male: A patient with huge maxillary Squamous Cell Carcinoma infiltrating soft tissues on all sides underwent 6 cycles chemotherapy with 5-FU, Cisplatin and Docetaxel with reasonable regression. He was operatedand kept on well for about 2 months only when the tumor recurred rather aggressively. Palliation was attempted with paclitaxel and the tumor responded to an extent but myelosuppression was noted with both leuco- and thrombocytopenia. This was managed along with corticosteroids & antibiotics and G-CSF. After whole blood transfusions and at least 7 platelet rich transfusions, Leucopenia responded but thrombocytopenia persisted around 50,000 per mm3. It improved with administration of Platelet Boost capsules. When further chemotherapy was administered, his condition became rather low and he could no longer swallow the platelet medicine. A special liquid form was made for him and his thrombocytes remained at a platelet around 100000/c.mm. 6. MH: 13 years. A young boy presented with Osteosarcoma of left lower femer and it was decided to give him an aggressive course of 6 cycles of neoadjuvant chemotherapy with high doses of Doxorubicin, Ifosfamide, Cisplatin and Etoposide. With the first cycle, his degree of myelosuppression was severe with leucopenia 750/mm³ and thrombocytopenia with 44000/mm³ and he improved rapidly with the usual treatment for leucopenia and Platelet Boost Capsules. He is now continuing the PB treatment prophylactically for the subsequent cycles of chemotherapy with no further thrombocytopenia.

Claims

1. A pharmaceutical composition for treating thrombocytopenia, which pharmaceutical composition comprises a pharmaceutically acceptable carrier and an active compound composition comprising one or more compounds present in papaya leaf dry matter chosen from the group consisting of papain, chymopapain, alkaloids, benzylglucosinolate and tannins.

2. A pharmaceutical composition according to claim 1, wherein said alkaloids are chosen from the group consisting of carpaine, nicotine, myosmine, piperidine, dehydrocarpaines, pseudocarpaine.

3. A pharmaceutical composition according to claims 1 or 2, wherein said active compound composition is papaya leaf dry matter.

4. Method for treating a mammal, in particular a human, with a deficiency in platelets by administration of a pharmaceutical composition according to claims 1 - 3.

5. Method according to claim 4, wherein said pharmaceutical composition is papaya leaf dry matter and wherein said papaya leaf dry matter is preferably administered in a daily dosage of at least 50 mg, more preferably a daily dosage of at least 100 mg.

6. Method according to claim 5, wherein said daily dosage is divided over smaller dosages administered to the patient over the course of the day.

7. Method according to any of claims 4 - 6, wherein said pharmaceutical composition is administered orally.

8. Method according to any of claims 4 - 7, wherein said papaya leaf dry matter is administered in a daily dosage of 2-3 mg/kg, based on the bodyweight of said mammal.

9. Method according to any of claims 4 - 8, wherein said mammal is receiving chemotherapy treatment.

10. Use of a pharmaceutical composition according to any of claims 1 - 3 as a prophylactic, in particular for administration in a patient receiving chemotherapy.

11. Method for preparing a pharmaceutical composition according to any of claims 1 - 3, comprising the steps of:

- providing papaya leaves, preferably fresh matured papaya leaves; and

- drying said papaya leaves, preferably by freeze drying, thus obtaining papaya leaf dry matter; and

- encapsulating the papaya leaf dry matter into a pharmaceutically acceptable carrier.

12. Method according to claim 11, wherein the capsule obtained after encapsulating said granules comprises 10 - 2000 mg of said granules, preferably 80 - 500 mg of said granules, more preferably 150 - 350 mg of said granules.