RU2411935C1 - Pharmaceutical composition based on doxorubicine and phospholipid nanoparticles for treatment of oncologic diseases - Google Patents

Abstract

FIELD: medicine.

SUBSTANCE: invention relates to field of medicine, in particular to pharmaceutical composition for treatment of oncologic diseases in form of phospholipid nanoparticles with size 10-30 nm, which includes phosphatidelcholin, maltose and doxorubicine with the following ratio of components, wt. %: phosphatidelcholin 20-43, maltose 55-78, doxorubicine 2-8. Composition is accumulates in tumour tissue more actively and slows down tumour growth in mice with carcinoma LLC more efficiently in comparison with free doxorubicine.

EFFECT: composition represents freeze-dries powder, stable in long storage, which dissolving in water gives nanophpospholipid particles with included doxorubicine.

3 dwg, 5 tbl

Description

The invention relates to medicine and pharmacology, and relates to a storage stable and effective in its specific action drug composition, which consists of nanoparticles based on plant phospholipids, including the antitumor drug doxorubicin.

Doxorubicin is a well-known antitumor antibiotic of the anthracycline series, its chemical name is (8S-cis) -10- (3-amino-2,3,6-tridesoxy-alpha-L-lyxohexopyranosyl) oxy-7,8,9,10- tetra-hydro-6,8,11-trihydroxy-8- (hydroxylacetyl) -1-methoxy-5,12-naphthacenedione, gross formula C ₂₇ H ₂₉ NO ₁ . The doxorubicin molecule consists of a tetracyclic anthraquinoid aglycon of doxorubicinone, coupled by a glycosidic bond with aminosugar daunosamine

For more than 30 years, doxorubicin has been used in the treatment of both hematological oncological diseases and solid tumors of various localization [1, 2].

Its cytostatic effect is due to the binding of tumor cells to DNA, inhibiting their growth and proliferation [3]. The drug can be introduced between the layers of DNA base pairs, inhibiting DNA-dependent RNA synthesis.

Doxorubicin is used only in the form of solutions for intravenous and intravesical administration. Intravenously, it is administered at 60-75 mg / m ² once every 3-4 weeks, or 30 mg / m ² once a week for 3-4 weeks. After iv administration, doxorubicin quickly disappears from the blood, distributed to organs and tissues of the body: kidneys, myocardium, spleen, lung. In the liver, it is metabolized to form the active metabolite of doxorubicinol. The half-life of doxorubicin is 20-48 hours, while 40% of it is excreted in the bile unchanged for 7 days, 5-12% of doxorubicin and its metabolites are excreted in the urine within 5 days.

A limitation for the therapeutic use of doxorubicin is its toxicity to healthy tissues, primarily cardiotoxicity, which manifests itself in severe heart failure. An effective way to reduce it is considered to be the use of doxorubicin not in free form, but as part of medicinal compositions: doxorubicin and a carrier (transport system), which affect the biodistribution of the drug in the body and, in particular, reduce the possibility of its entry into the heart [4].

In this regard, in recent years, a large number of research and development of new dosage forms of doxorubicin equipped with a delivery system in the body has been devoted to doxorubicin in particular.

The main requirement for such forms of doxorubicin is to maintain or increase the therapeutic antitumor effect while reducing side effects. The most common delivery systems are liposome forms based on lipids [4-6]. The advantages of the liposomal form are currently well known and are associated primarily with improved pharmacokinetics and reduced side effects. For doxorubicin, this form is very productive, since a drug encapsulated in liposomes practically does not enter healthy tissues, penetrating only through defective capillaries of tumors, and the amphiphilic phospholipid membrane of liposomes, interacting with the cell membrane, can facilitate the interaction of the drug with the target cell, thereby increasing its very effect. In this regard, the size of phospholipid carriers is significant - its maximum reduction should facilitate penetration into the tumor, including intracellular drug delivery, and also increase resistance to particle capture by the reticuloendothelial system (RES).

Several liposomal forms of doxorubicin are known with an average diameter of 100 to 400 nm, which are two groups of drugs. The most commonly used drug is doxyl (doxyl), also known as kelix (caelyx), manufactured in the USA [6]. The drug is approved for use in Russia. It is a stabilized polyethylene glycol (PEG) liposome containing doxorubicin. To obtain them, a mixture of lipids (hydrogenated soybean phosphatidylcholine, cholesterol and distearoylphosphatidylethanolamine with attached PEG in a ratio of 56: 39: 5) is suspended in a solution of 250 mM ammonium sulfate, and then subjected to extrusion (i.e., homogenization with filtration under pressure). Unencapsulated ammonium sulfate is removed (for example, using column chromatography), doxorubicin is added and incubated at 55-60 ° C. In this case, due to the pH gradient, the drug enters the liposome, forming a salt with a captured sulfate anion, which stabilizes it in the inner space of the liposomes. After the loading step, the mixture is cooled. According to the same principle, but using mainly egg phosphatidylcholine, the drug lipodox was designed (Ukraine) [7]. At the same time, the presence of PEG in such preparations can lead to additional side interactions.

Another form of PEG free liposomal doxorubicin is also known. This is Miocet (TLC D-99, manufactured by The Lyposome Company, USA), which is phosphatidylcholine cholesterol liposomes containing doxorubicin citrate with a drug: lipid ratio of 0.28 [8]. To obtain it, first prepare single-layer liposomes of an average diameter of 180 nm from a mixture of egg phosphatidylcholine with cholesterol (55:45) in citrate buffer and incubate them with shaking with a freshly prepared solution of doxorubicin chloride with methyl paraben at 55-60 ° C. The disadvantage of this unstabilized form is the rapid absorption of liposomes by RES cells, which is facilitated by their relatively large size - above 150 nm, in combination with an unprotected surface.

Thus, each of the liposomal forms of doxorubicin available on the market has certain disadvantages: low stability or additional side effects. In connection with the foregoing, the search and development of its new optimized lipid dosage forms is currently ongoing. So, doxorubicin was embedded in phospholipid nanoparticles that make up the injectable form of the phosphogliv drug with a particle size of ~ 50 nm [9]. For this, a weighed portion of doxorubicin (100 mg) was added to the mixture of the starting substances of the phosphogliv preparation: 5 g of soy phosphatidylcholine (Lipoid S100, Lipoid, Germany), 2 g of glycyrrhizic acid trisodium salt (China) and 20 g of maltose monohydrate. 100 ml of water for injection was added, mixed on a vibrating mixer and the resulting aqueous suspension was homogenized using a high pressure homogenizer RANNIE MiniLab 7.30 VH (Denmark) for 5 minutes. The resulting solution was subjected to sterilizing filtration using filters with a pore diameter of 0.22 μm with an excess nitrogen pressure of 1.5-2 atm, poured into sterile 10 ml vials and lyophilized using freeze-drying Liolab F. The drug showed more pronounced antitumor and antimetastatic activity in mice with LLC carcinoma compared to free doxorubicin. Also described are liposomes with a diameter of ~ 135 nm from phosphatidylcholine, cholesterol and oleic acid, with 0.19 moles of doxorubicin per mole of lipid obtained by injection of an ethanol solution [10], solid lipid nanoparticles from monostearin, octadecylamine and oleic acid [11], heat-sensitive liposomes [12]. Targeted liposomes with doxorubicin were also developed, containing estrone [13] or interleukin-13 [14] as a ligand — increased expression of receptors, which, according to some data, is manifested on a number of tumor cells. The preparation of “nanoassemblies” of phosphatidylethanolamine with PEG with a size of 10–20 nm, which exhibit a pronounced therapeutic effect on the model of LLC carcinoma in mice [15], is also described. At the same time, it is impossible not to mention that none of these works paid attention to the intracellular penetration of doxorubicin introduced in new transport forms, its interaction with DNA, which is the basis of the cytostatic effect, and acquires special significance when passing to nanoscale particles that can deliver medicine not only to the cell membrane, but also penetrate the cell.

The objective of the present invention is to develop a non-fat phospholipid composition of doxorubicin with an average nanoparticle diameter of 10-30 nm, which has reduced toxicity compared to a free drug while maintaining a specific cytostatic effect, capable of withstanding long-term storage and transport of doxorubicin into the tumor tissue, ensuring its high bioavailability.

The problem is solved by creating a pharmaceutical composition for the treatment of cancer in the form of phospholipid nanoparticles with a size of 10-30 nm, including plant-derived phosphatidylcholine, maltose and doxorubicin in the following ratio, wt.%:

Phosphatidylcholine 20-43 Maltose 55-78 Doxorubicin 2-8

Used phosphatidylcholine is the main component of highly purified vegetable soybean phospholipid with a content of at least 73-95% wt. Other phospholipid components may be contained in amounts not exceeding permissible (lysophosphatidylcholine up to 4% wt., Trace amounts of other phospholipids).

As auxiliary pharmacologically acceptable substances, the composition contains maltose, which is a cryoprotectant at the stage of freeze-drying. The addition of maltose makes it possible to obtain a lyophilisate capable of completely recovering its structure after rehydration, in particular particle size.

According to the invention, the proposed composition allows to obtain a pharmaceutical preparation with a large accumulation of doxorubicin in the tumor tissue compared to the original drug.

A method of obtaining a phospholipid composition of doxorubicin is as follows.

Materials and methods

The following materials were used in the work:

1. Soy phospholipid company Lipoid GmbH, Germany, with a phosphatidylcholine content of 78-95%.

2. Maltose monohydrate company MERCK, Germany.

3. Water for injection (according to FS No. 42-4587-95).

4. Doxorubicin, substance (according to FS 42-02402570-02).

Example 1. Obtaining doxorubicin in the composition of phospholipid particles

A. Obtaining a coarse emulsion

25 g of maltose are dissolved in 200 ml of water with a temperature of 45 ° C (until complete dissolution). 625 mg of doxorubicin and 6.25 g of phospholipid are added to the resulting maltose solution. Using a household blender, homogenize the resulting mixture and bring its volume to 250 ml.

B. Obtaining Doxorubicin in the composition of phospholipid nanoparticles

The resulting coarse emulsion is passed through a homogenizer (MiniLab 7.3 VH, Rannie, Denmark) at a pressure of 800 bar. Light transmission is recorded at 660 nm. Filter the drug through a 0.22 micron filter. Re-register light transmission at 660 nm, observe its increase. The particle size in the preparation is determined. The doxorubicin content in the resulting preparation is analyzed by high performance liquid chromatography (HPLC). The drug is poured into 10 ml vials and freeze-dried.

The contents of the vial are dissolved in 10 ml of purified water and the particle size is determined after dissolving the freeze-dried powder of the drug. The particle size is determined in a day, two and a week of storage of the dissolved drug at a temperature of 4 ° C.

Characterization of the composition of doxorubicin in the composition of phospholipid nanoparticles

Particle size analysis using a Beckman N5 device (see table 1) indicates that more than 96% of the particles have a size of (20.0 ± 2.0) nm. The incorporation of doxorubicin into phospholipid nanoparticles is about 96%.

Table 1 Particle size distribution in doxorubicin in nanophospholipid particles N measurements Range of measurement, nm Particle distribution size, nm content,% standard deviation, nm one 3.0-450.0 19.5 95.17 1.3 2 3.0-450.0 21.3 96.50 1,2 3 3.0-450.0 21.1 96.34 2.1 four 3.0-450.0 20.6 95.97 1,6 5 3.0-450.0 19.7 97.21 1.4

The determination of the drug substance in the phospholipid preparation was carried out by HPLC. For this, a 9-fold excess of methanol was added to the preparation, thoroughly mixing on a mixer. HPLC analysis of aliquots of the prepared solution (1-10 μl) was performed on an Agilent Technology 1200 (USA) or Milichrom (Novosibirsk) instrument using a linear gradient of acetonitrile in a 0.1% aqueous solution of TFU, from 1% to 60%, at a rate elution of 200 μl / min and detection in the range of 220-360 nm. To determine the concentration, we used a calibration curve of the dependence of the peak area on the number of corresponding standard samples of substances.

According to HPLC, the incorporation of doxorubicin into phospholipid nanoparticles is about 96%. After rehydration of the lyophilized dried doxorubicin preparation as part of phospholipid nanoparticles (NF-doxorubicin), the nature of the particle size distribution and the size of the main particle size fraction does not change: more than 96% of the particles have a size of (20.0 ± 2.0) nm. When storing the dissolved drug at 4 ° C for 7 days, the particle size is preserved.

table 2 Particle size distribution in the solution of the drug NF doxorubicin at the time of release, after 4 and 7 days of storage N measurements Shelf life Range of measurement, nm Particle distribution size, nm content,% standard deviation, nm one 2 3 four 5 6 in the moment 3.0-450.0 one release 19.5 96.33 1.3 2 18.9 97.13 1,6 3 19.6 96.43 1.9 four 20.1 96.87 2.1 4 days 3.0-450.0 one 20.3 97.26 1,6 2 19.5 96.37 1.7 3 19,4 96.96 2.0 four 19.6 96.53 1.8 7 days 3.0-450.0 one 19.8 96.21 1.7 2 20,4 96.15 1.9 3 19.7 95.98 2,3 four 20.9 95.73 1.4

According to the invention, the proposed composition allows to obtain a pharmaceutical preparation with greater availability of doxorubicin in relation to tumor tissue compared to the free drug.

To evaluate the effect of the obtained drug composition of doxorubicin in experimental animals, its accumulation in the tumor and distribution by blood and plasma fractions, it was necessary at the first stage to develop a method for extracting doxorubicin from tissues, which would allow monitoring not only free drug, but associated with cells in in particular as part of complexes with DNA.

List of drawings

Figure 1 - binding to red blood cells of free doxorubicin and doxorubicin in the composition of phospholipid nanoparticles during incubation in vitro with heparinized blood.

Figure 2 - distribution between the lipoprotein fractions in the blood plasma of free doxorubicin and doxorubicin in the phospholipid nanoparticles after incubation of the plasma in vitro with blood plasma.

Figure 3 - release of Triton X-100 doxorubicin (detected after ultrafiltration) in plasma incubated with free or with NF-doxorubicin.

The choice of doxorubicin extraction method

A method was selected for the extraction of doxorubicin with a mixture of methanol and 0.5% trifluoroacetic acid (TFA) and the completeness of the extraction of the preparation from complexes with DNA was shown (Table 3). For this, the following procedure was performed:

- to 100 μl of a DNA solution (10 μg / ml) was added 100 μl of a pre-diluted (1: 100) solution of doxorubicin;

- 50 μl of a solution of the doxorubicin-DNA complex was taken and 450 μl of extractant was added: a) methanol and b) methanol with 0.5% TFA (i.e., the concentration of total doxorubicin in each sample was 1000 ng / ml);

- the solutions were mixed on a vibrator, centrifuged, and 20 μl was taken from each solution for analysis of doxorubicin by HPLC, while in each sample the amount of doxorubicin corresponded to 10 ng of the drug in its complex with DNA.

Table 3 The completeness of the extraction of doxorubicin from complexes with DNA Extraction method The detected amount of doxorubicin in the extract (in 20 μl), ng The initial amount of doxorubicin in the sample, ng The percentage extraction of doxorubicin from complex with DNA one 2 3 four Methanol one 10 10% Methanol with 0.5% TFA 10 10 one hundred%

Since treatment with a mixture of methanol with 0.5% TFA leads to the complete extraction of doxorubicin, this system was chosen for the subsequent extraction of doxorubicin from tissues.

The effect of free doxorubicin and doxorubicin in the composition of phospholipid nanoparticles in mice with an inoculated tumor

Comparison of drug accumulation in tumor tissue and in the liver

Experiment

Used mice with an inoculated Lewis LLC adenocarcinoma.

Tumor volume ~ 5000 mm ³

Weight of mice ~ 20-25 g

Doxorubicin preparations were administered intraperitoneally to mice at a dose of 15 mg / kg body weight.

After 4 hours, the animals were killed, the liver and tumor were removed.

Tissue samples were homogenized in a mixture of methanol with 0.5% TFA in the ratio of 100 mg of tissue per 900 μl of solvent. The homogenate was centrifuged; in the supernatant, the concentration of doxorubicin was determined by HPLC. The results are shown in table 4.

Table 4 The content of doxorubicin in the liver and tumor of mouse tumors after administration of free doxorubicin and NF-doxorubicin the cloth The amount of doxorubicin, mcg / g of tissue after administration of free doxorubicin after administration of NF-doxorubicin Liver 2.5 3,1 Tumor 1,2 5,4

From the above data it follows that in the case of using NF-doxorubicin, 4.5 times more medicine enters the tumor (target tissue) than when administered in free form.

Comparison of the specific antitumor activity of free doxorubicin and doxorubicin in the composition of phospholipid nanoparticles

Solutions of both forms of doxorubicin were intraperitoneally administered to mice with LLC tumor at a dose of 5 mg / kg. The drugs were administered, starting from 7 days after tumor transplantation, once a week (i.e., only three injections). Tumor sizes were determined 3 and 15 days after the start of treatment. The results of the experiment are given in table 5.

Table 5 Inhibition of tumor growth with the administration of doxorubicin and NF-doxorubicin to mice Groups n The average volume of tumors, mm ³ Day after starting treatment 3 days after the 1st injection After 15 days (i.e. after the 3rd injection) The control 13 1156 [769 ÷ 1543] 6713 [5082 ÷ 8344] Oxoxorubicin 10 834 [541 ÷ 1127] 5390 [4161 ÷ 6619] Tumor growth inhibition,% 28 twenty NF-doxorubicin 10 349 [199 ÷ 499] 2934 [2358 ÷ 3510] Inhibition of tumor growth,% 70 * 56 * * - significantly with respect to the group of animals that received the free drug, p <0.05.

When administered in this mode, NF-doxorubicin causes a significantly more pronounced inhibition of tumor growth compared to free drug.

Thus, doxorubicin in the composition of phospholipid nanoparticles has a higher antitumor activity in the experimental tumor model as compared to free doxorubicin, which was manifested in increased accumulation of the drug in the tumor tissue and more inhibition of tumor growth.

To characterize the properties of doxorubicin in the composition of phospholipid nanoparticles, which may affect its specific activity, in vitro experiments were carried out to incubate NF-doxorubicin with heparinized blood taken from male Wistar rats weighing about 400 g. The distribution of the drug in the blood was examined both between red blood cells and plasma, and in plasma between separate classes of lipoproteins in healthy animals.

Distribution of free doxorubicin and doxorubicin in the composition of phospholipid nanoparticles in the blood

Distribution between blood cells and plasma

There is literature evidence that doxorubicin, due to its high nonspecific absorption activity, binds to a large extent with red blood cells when introduced into the blood [16]. This reduces the proportion of active drug in plasma, weakening the total therapeutic effect. Therefore, when developing the NF-doxorubicin composition, it seemed appropriate to evaluate the possible change in such non-specific binding in a new dosage form.

The experiment:

1) 100 μl of a solution of doxorubicin or NF-doxorubicin with a concentration of 2 mg / ml (which corresponds to a concentration of the drug in a blood sample of 200 μg / ml) was added to 900 μl of heparinized blood;

2) incubated at 37 ° C with stirring for 30 or 60 minutes;

3) 50 μl of blood was taken, 450 μl of a mixture of methanol with 0.5% TFA was added, mixed on a vibrator, centrifuged and the supernatant was taken for analysis;

4) 100 μl of blood from the same tube was centrifuged to precipitate the shaped elements. 450 μl of a mixture of methanol with 0.5% TFA were added to 50 μl of plasma, mixed on a vibrator and centrifuged, selecting the supernatant for analysis;

5) in the obtained acid-methanol extracts from whole blood and plasma, the content of doxorubicin was determined by HPLC. The results were counted on the drug content in 1 ml of the original whole blood (figure 1). The content of doxorubicin in the formed elements was determined by difference.

In the drawing, it can be seen that when doxorubicin is included in the composition of phospholipid nanoparticles (NF-doxorubicin), the percentage of the drug associated with the formed elements of the blood is reduced. This increases the relative proportion of free drug that can have a therapeutic effect.

Plasma doxorubicin distribution - for free drug and NF-doxorubicin

In recent years, the effect on the pharmacokinetics and distribution of the drug in the body of the nature of its distribution in blood plasma, i.e. binding to individual classes of lipoproteins and plasma proteins (primarily with albumin). Since 2002, the US Food and Drug Administration (FDA) has made this study mandatory when registering new drugs that include lipophilic components [17]. Therefore, it seemed appropriate to characterize in this aspect NF-doxorubicin.

The experiment:

1) rat blood was collected in a test tube with heparin (8 ml of blood per 1.6 ml of heparin solution), centrifuged, and 3.8 ml of plasma were collected in two tubes;

2) 200 μl of a solution of doxorubicin or NF-doxorubicin was added to a plasma concentration of 0.1 mg / ml;

3) incubated for 30 minutes at 37 ° C;

4) in order to fractionate lipoproteins by densities, 2 g of crystalline NaBr was added, mixed until the salt was completely dissolved and 6 ml of a NaBr solution with a density of 1.019 g / ml was layered;

5) centrifuged on an Optima L90K Beckman ultracentrifuge at 41,000 rpm in an angular rotor of 65 Tu at 4 ° C for 2 hours;

6) fractions were collected from above the height of the tube, 1-2 ml, corresponding to very low lipoproteins (VLDL), low (LDL) and high density and the remaining bottom protein fraction (without lipoproteins);

7) aliquots of each fraction were treated with a 9-fold volume of methanol, the protein precipitate was separated by centrifugation (10 min at 3000 rpm), and the methanol extracts were analyzed by HPLC. The results are presented in figure 2.

From the presented drawing it is seen that when using NF-doxorubicin, the distribution of the drug shifts from the albumin fraction of the plasma to lipoproteins, mainly to HDL, which may also be partially responsible for its greater penetration into the tumor tissue.

Assessment of the relative binding strength of doxorubicin in the composition of nanoparticles of the NF-doxorubicin composition

An additional characteristic of the doxorubicin drug composition is also the duration of drug retention in the nanophospholipid particle. A criterion for this could be the degree of binding of the free drug to plasma proteins - in the case of plasma incubation with free or with NF-doxorubicin. To detect this binding, incubated plasma ultrafiltration was used in combination with treatment with Triton X-100, which destroys protein complexes, with the release of free drug.

The experiment:

1) free or NF-doxorubicin was added to 900 μl of plasma to a concentration of doxorubicin 0.5 mg / ml and incubated for 4 hours at room temperature;

2) Triton X-100 was added to a concentration of 1% and incubated for 10, 30 or 90 minutes;

3) the plasma after incubation with Triton X-100 was ultrafiltered, the ultrafiltrates were treated with methanol as described above, and the concentration of doxorubicin was determined in methanol extracts by HPLC. The results are presented in figure 3.

Due to the fact that during ultrafiltration of plasma, proteins are retained and do not pass into the filtrate, the absence of doxorubicin in it indicates the absence of free drug in the plasma, in both cases. In the case of plasma with free doxorubicin, its appearance during treatment with triton, up to an almost complete yield, indicates that doxorubicin was initially in complex with plasma components that are destroyed by triton. In a plasma incubated with NF-doxorubicin, the release of free drug occurs 1.5 times slower. Thus, we can conclude that a significant part of the drug during the 4-hour incubation was protected from complexation with plasma proteins, since it was part of a nanophospholipid particle.

Indicators characterizing the degree of toxicity of free doxorubicin and doxorubicin in the composition of phospholipid nanoparticles

A comparative assessment of the toxicity of both forms of doxorubicin was carried out on male BDF1 mice weighing at least 18-20 g. The LD ₅₀ value was determined with a single intravenous administration:

LD ₅₀ for NF-doxorubicin - 12.4 mg / kg;

LD ₅₀ for the drug free doxorubicin (LENS Pharm) - 12.8 mg / kg.

Treatment tolerance was also comparable: no deaths were observed in any groups.

Thus, doxorubicin in the phospholipid composition in the form of nanoparticles of 10-30 nm accumulates more actively in the tumor tissue and inhibits tumor growth more effectively in mice with LLC adenocarcinoma. In vitro experiments for this composition (tentatively called “doxolip”) showed a large proportion of doxorubicin not binding to blood cells, as well as its redistribution in blood plasma from the protein fraction to lipoproteins. The stability of doxolip particles during incubation with blood for 4 hours is shown. In terms of toxicity, doxolip is comparable to free doxorubicin produced by LENS Pharm.

LITERATURE

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2. The register of medicines of Russia, M., 2000

3. Terasaki T., Iga T., Sugiyama Y., Sawada Y., Hanano M. Nuclear binding as a determinant of tissue distribution of adriamycin, daunomycin, adriamycinol, daunorubicinol and actinomycin D. J. Pharmacobiodyn. 1984, 7 (5), 269-277.

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8. Cowens J.W., Creaven P.J., Greco W.R., Brenner D.E. Doxorubicin Encapsulated in Liposomes. Initial Clinical (Phase I) Trial of TLC D-99. Cancer Research 1993, 53, 2796-2802.

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Claims (1)

A pharmaceutical composition for the treatment of cancer in the form of phospholipid nanoparticles with a size of 10-30 nm, including 78-95% plant-based phosphatidylcholine, maltose and doxorubicin in the following ratio, wt.%:
Phosphatidylcholine 20-43 Maltose 55-78 Doxorubicin 2-8

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