Dear collegues and patients,
I'd like to present my pharmaceutical experience.
I'm unable t post pictures, though, so if you want to view them then e-mail me.
Regards,
Professor, Academician NAN RK
Murat Gilmanov
PHOSPHATIDYLINOSITOL NANOCONTEINERS LOADED WITH MEDICINE FOR TREATMENT OF CARDIO, ENDOCRINE AND INFECTIOUS DISEASES
M. K. Gilmanov. S. M. Gilmanova
(phone/fax: +7 727 394 6274; mobile : +7-705-622-4011 ; e-mail: baltakay@mail.ru).
Paper is devoted to the development of a new drug delivery system for universal medical application. This drug delivery system represents unique nanoconteiners that consist of phosphatidylinositol (PI) molecules. We have developed the new effective methods of preparation of PI from plant organelles - spherosomes. Spherosomes were purified using a novel nanostructured carbon sorbent – “nanocarbosorb”. Also we have worked out methods of preparation and loading of nanoconteiners. The nanoconteiners loaded by drugs were tested for treatment of cardio, endocrine and infectious diseases like heart attack, diabetic foot, glaucoma, arthritis and tuberculosis in several medical scientific institutes of Almaty. As nanoconteiners consist of natural phosphatidylinositol they easily penetrate the cells and leave no remains, as they will be used in the synthesis of cell membranes, whereas, synthetic drugs are very difficult to enter the cells of affected organs. Moreover, nanoconteiners allow the reduction of the dose of the medicine by 100 times during the treatment course and consequently the toxic effect on the whole organism also reduces by 100 times. Thus, drug loaded nanoconteiners allow the effective treatment of various diseases without toxic damaging effect on other organs. Altogether it gives good medical and economical effects.
The problem of medicaments delivery to the affected organs has no satisfactory solution up to the present days. All existing drug delivery systems have some serious disadvantages or other. So lecithin liposomes are too large, unstable and might easily aggregate into big complexes. This causes a danger of vascular occlusion. The nanoconteiners made from plastics causes immune and allergic reactions to the patient, which is also a big disadvantage. For this reason one of the main problems of modern medicine is to develop new generation of drug delivering systems. Here we would like to present our scientific investigations that by far lead to the development of innovative technologies in the field of nanotechnology.
In the Laboratory of Enzyme Structure and Regulation of Aytkhozhin’s Institute of Molecular Biology and Biochemistry of the Republic of Kazakhstan for the first time we have determined the structure of plant subcellular organelle - spherosome. Now it is clear that spherosomes consist of only one phospholipid – phosphatidylinositol (PI) and only one protein – glutamate dehydrogenase (GDh) [1].
The unique structure of spherosome allows the development of the new technology. Since spherosomes contain only one type of phospholipid – PI, it is very convenient to develop the method of isolation of chromatographically pure PI fractions. This method is protected by US patent ? 4,977,091 [2]. Therefore, we issue a challenge to create drug delivery systems on the basis of PI. Another task of our investigation was the development of the new effective, but simple methods to isolate spherosomes using the nanostructured carbon sorbents.
The price of purified preparations of PI at the market was very high before our method was developed. For example, in 2006-2007 Sigma catalogue the price of PI preparation (catalogue number PO639-50MG) is 641$ per 50 mg, whereas, our method allows to reduce this price by 10-100 times for the same product. Thus, our method opens a new way for wide industrial application of PI.
First of all we attempted to create PI liposomes on the basis of our PI product. As our endeavors we successful we have obtained PI liposomes that have sizes less than 1 ?m and undoubtedly can be called nanoconteiners. These nanoconteiners are found to be very stable for months. Negatively charged our PI nanoconteiners push off each other and so will never form aggregates in the patient’s organism. We have also developed new effective method to load our PI nanoconteiners with medicine. We used the property of PI nanoconteiners to open in the hydrophobic solution as it is shown in Fig. 1.
[/img]Fig. 1. Electronic microscopy of opening nanoconteiners after transferring them into hydrophobic solution (95% ethanol).
The loading procedure is very simple. First of all PI nanoconteiners must be treated with ethanol solution that allows them to open like shells. Then PI nanoconteiners are transferred into usual buffer solution containing drug molecules where they begin to close. During this process drug molecules enter the inner part of PI nanoconteiners. This method is protected by the patent of the Republic of Kazakhstan.
The nanoconteiners loaded by piroxicam, diclofenac, isosorbide dinitrate, rifampicine, enambutol, pyrasinamide and vasoprostan were tested for treatment of diseases in several medical scientific institutes of Almaty.
Nanoconteiners loaded by cardio drugs were tested in the Institute of Cardiology and Internal Diseases of Ministry of Healthcare of the Republic of Kazakhstan under the supervision of professor Ablayuly and doctor Mansharipova. Our nanoconteiners were loaded by isosorbide dinitrate, pharmaceutical trade name is “Isoket”. This drug is normally used for heart attack prevention and its mechanism of action is the same as nitroglycerol. We have prepared the ointment consisting of “Isoket” loaded nanoconteiners and lanolin. The experiments were carried out on rats with artificial infarct model. Induced heart attack in rats was successfully prevented by applying the prepared ointment onto the skin of rat chest in heart area. The heart attack was stopped in 10-15 minutes after the ointment was rubbed into the chest of experimental rat. The improvement of heart action was registered by the increase of oxygen supply to the heart muscles and blood vessels. The nitrogen oxide signal induced by ”Isoket” is very important in the heart oxygen supply. The content of NO was determined and the results are given in table 1. Nowadays, the nanoconteiners are being tested on volunteers in this institution.
Groups/ samples Control group Experimental group before nanoconteiner treatment Experimental group after nanoconteiner treatment
Coronary vessels 16,18±1,35 6,14±0,52* 18,92±1,31°
Myocardium tissues 14,25±1,96 3,11±0,24* 32,93±1,91°**
Table 1 – the content of NO in different tissues before and after “Isoket” loaded nanoconteiner treatment (nmol/ml)
Moreover, the nanoconteiners loaded by prostaglandin vasoprostan with strong vasodilatory activity have been generated in our laboratory and now are being applied to the treatment of the diseases like diabetic foot and eye diseases. They were also tested in the Institute of Eye Diseases (Almaty, Kazakhstan) under the supervision of Dr. Dzhumatayeva Z.A. The tests showed that our nanoconteiners loaded by vasoprostan show good therapeutic effect in the treatment of glaucoma.
On the basis of nanoconteiners we have created new anti-arthritic drugs. Nanoconteiners were loaded with non-steroidal anti-inflammatory drugs – piroxicam or diclofenac. They were tested in rheumatological centre of Almaty on experimental and control groups of arthritic patients under supervision of Professor Seisenbayev A.S. Each group comprised of thirty patients comparable by age, sex and stage and activity of the disease. The ointment containing our loaded PI nanoconteiners were applied for treatment of the patients of experimental group. After 3 weeks there was a significant remission of a local painful syndrome in 76.6% of patients in experimental group and only 50 % - in control group, which had taken a traditional treatment. Thus the obtained anti-inflammatory preparation on the basis of PI nanoconteiners shows the better treatment results than standard drugs. Our preparation has next advantages in comparison with standard drugs:
- the reduction of therapeutic doses of piroxicam or diclofenac more than ten times with the same or better therapeutic effect;
- ability to penetrate deeper and faster into the tissues of joints than standard drugs and prolongation of the action of medicine;
Fantastic results are achieved in the treatment of tuberculosis of spinal cord spondyls with nanoconteiners. The experiment was carried out in the Kazakh Scientific Research Institute of Tuberculosis by Doctor Tutkishbayev S.O. Four different types of nanoconteiners loaded with rifampicine, enambutol, pyrasinamide and isoniaside were prepared at a time. The ointments were applied to the affected spondyls one after another every morning and evening during 40 days and as a result the disease was completely cured in only one month time, whereas the traditional treatment require 4-5 months of intensive therapy.
Thus we have developed the new generation of drug delivery system namely PI nanoconteiners. These nanoconteiners have obvious advantages than the traditional treatment, which uses tablets and/or injections. The distinctive features of nanoconteiners are very small sizes and thermal movement that allows them to reach the affected organ in approximately 15 min. As nanoconteiners consist of natural phosphatidylinositol they easily penetrate the cells and leave no remains as they will be used in the synthesis of cell membranes, whereas, synthetic drugs are very difficult to enter the cells of affected organs. Moreover, nanoconteiners allow the reduction of the dose of the medicine by 100 times during the treatment course and consequently the toxic effect on the whole organism also reduces by 100 times. Thus, drug loaded nanocapsules allow the effective treatment of various diseases without toxic damaging effect on other organs. Also we have achieved the significant prolongation of the action of the medicine. Altogether it gives good medical and economical effects.
REFERENCES
[1] M. K. Gilmanov, R.Dilbarkanova, Structure and functions of plant cells spherosomes. Almaty: Gylym, 1997, pp. 52-88. (In russian)
[2] M.K Gilmanov, R. Dilbarcanova, B.E. Sultanbaev. // The method for preparing phosphatidylinositol from vegetable material // The Commissioner of patents and trademarks. Patent
? 4,977,091 USA, December 11, 1990.
