WO2022119531A1 - Investigation of the potential of pemetrexed-loaded liposome system in cancer treatment - Google Patents
Investigation of the potential of pemetrexed-loaded liposome system in cancer treatment Download PDFInfo
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- WO2022119531A1 WO2022119531A1 PCT/TR2021/051212 TR2021051212W WO2022119531A1 WO 2022119531 A1 WO2022119531 A1 WO 2022119531A1 TR 2021051212 W TR2021051212 W TR 2021051212W WO 2022119531 A1 WO2022119531 A1 WO 2022119531A1
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- Prior art keywords
- liposomes
- pemetrexed
- liposome
- drug
- chitosan
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Links
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- 229960005079 pemetrexed Drugs 0.000 title claims abstract description 24
- QOFFJEBXNKRSPX-ZDUSSCGKSA-N pemetrexed Chemical compound C1=N[C]2NC(N)=NC(=O)C2=C1CCC1=CC=C(C(=O)N[C@@H](CCC(O)=O)C(O)=O)C=C1 QOFFJEBXNKRSPX-ZDUSSCGKSA-N 0.000 title claims abstract description 24
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- 238000011835 investigation Methods 0.000 title description 2
- 229920001661 Chitosan Polymers 0.000 claims abstract description 23
- 239000002246 antineoplastic agent Substances 0.000 claims abstract description 7
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- 239000003814 drug Substances 0.000 claims description 28
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- 238000011068 loading method Methods 0.000 claims description 6
- 229940042880 natural phospholipid Drugs 0.000 claims description 6
- 238000003786 synthesis reaction Methods 0.000 claims description 6
- 229920000642 polymer Polymers 0.000 claims description 4
- IIZPXYDJLKNOIY-JXPKJXOSSA-N 1-palmitoyl-2-arachidonoyl-sn-glycero-3-phosphocholine Chemical compound CCCCCCCCCCCCCCCC(=O)OC[C@H](COP([O-])(=O)OCC[N+](C)(C)C)OC(=O)CCC\C=C/C\C=C/C\C=C/C\C=C/CCCCC IIZPXYDJLKNOIY-JXPKJXOSSA-N 0.000 claims description 3
- 235000012000 cholesterol Nutrition 0.000 claims description 3
- 229940107161 cholesterol Drugs 0.000 claims description 3
- 239000003937 drug carrier Substances 0.000 claims description 3
- 229940067606 lecithin Drugs 0.000 claims description 3
- 235000010445 lecithin Nutrition 0.000 claims description 3
- 239000000787 lecithin Substances 0.000 claims description 3
- WTJKGGKOPKCXLL-RRHRGVEJSA-N phosphatidylcholine Chemical compound CCCCCCCCCCCCCCCC(=O)OC[C@H](COP([O-])(=O)OCC[N+](C)(C)C)OC(=O)CCCCCCCC=CCCCCCCCC WTJKGGKOPKCXLL-RRHRGVEJSA-N 0.000 claims description 3
- 229920005615 natural polymer Polymers 0.000 abstract description 5
- 150000002632 lipids Chemical class 0.000 description 9
- 210000004027 cell Anatomy 0.000 description 6
- 239000002202 Polyethylene glycol Substances 0.000 description 5
- 229920001223 polyethylene glycol Polymers 0.000 description 5
- 239000000243 solution Substances 0.000 description 5
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 4
- 125000000129 anionic group Chemical group 0.000 description 4
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- 206010005003 Bladder cancer Diseases 0.000 description 1
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- 208000006105 Uterine Cervical Neoplasms Diseases 0.000 description 1
- 238000010521 absorption reaction Methods 0.000 description 1
- 230000002378 acidificating effect Effects 0.000 description 1
- 239000008186 active pharmaceutical agent Substances 0.000 description 1
- BFNBIHQBYMNNAN-UHFFFAOYSA-N ammonium sulfate Chemical compound N.N.OS(O)(=O)=O BFNBIHQBYMNNAN-UHFFFAOYSA-N 0.000 description 1
- 229910052921 ammonium sulfate Inorganic materials 0.000 description 1
- 235000011130 ammonium sulphate Nutrition 0.000 description 1
- 230000003432 anti-folate effect Effects 0.000 description 1
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- 229940127074 antifolate Drugs 0.000 description 1
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- 239000012062 aqueous buffer Substances 0.000 description 1
- 210000000481 breast Anatomy 0.000 description 1
- VSGNNIFQASZAOI-UHFFFAOYSA-L calcium acetate Chemical compound [Ca+2].CC([O-])=O.CC([O-])=O VSGNNIFQASZAOI-UHFFFAOYSA-L 0.000 description 1
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- 230000000144 pharmacologic effect Effects 0.000 description 1
- NBIIXXVUZAFLBC-UHFFFAOYSA-K phosphate Chemical compound [O-]P([O-])([O-])=O NBIIXXVUZAFLBC-UHFFFAOYSA-K 0.000 description 1
- 239000010452 phosphate Substances 0.000 description 1
- 229920001282 polysaccharide Polymers 0.000 description 1
- 239000005017 polysaccharide Substances 0.000 description 1
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- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 1
Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K9/00—Medicinal preparations characterised by special physical form
- A61K9/10—Dispersions; Emulsions
- A61K9/127—Liposomes
- A61K9/1271—Non-conventional liposomes, e.g. PEGylated liposomes, liposomes coated with polymers
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P43/00—Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
Definitions
- the invention relates to a liposome loaded with pemetrexed having a field of use in the healthcare sector in general, used as an anti-cancer drug developed for use in cancer treatment in particular, and coated with natural polymer, chitosan.
- Cancer is one of the diseases that poses a great problem for all countries in the world today. Estimates show that 30 million people will die of cancer every year by 2030. Surgery, radiation, chemotherapy, and immunotherapy are the four main methods used for cancer treatment worldwide.
- Chemotherapy is one of the methods used in the treatment of cancer.
- high toxicity, poor oral bioavailability, low therapeutic indices, non-specific biodistribution, and the effect of an anticancer drug on both normal and cancerous cells are the main deficiencies of the chemotherapy method.
- Liposomes which are among the drug carrier systems, are used to eliminate or minimize the negative effects occurring in traditional treatment methods, to increase reaching cellular levels, to provide effective and reliable treatment at low dosages, to eliminate or minimize toxic effects and immunogenic factors, and to obtain the desired pharmacological response in the targeted area.
- Liposomes are used as a drug delivery system for different chemotherapies in the present art. Hydrophilic drugs are added to the aqueous phase during liposome synthesis, whereas lipophilic drugs are added to the organic phase. Drugs can be loaded into liposomes via active or passive loading methods. Drugs are first dissolved in water or in an organic phase; afterwards, drug-containing liposomes are prepared by appropriate methods in the passive loading method. Active drug loading means that drugs are loaded into preformed liposomes. The drug can be actively loaded into the aqueous nucleus of a liposome by means of a chemical gradient, such as pH, ammonium sulfate, or calcium acetate.
- a chemical gradient such as pH, ammonium sulfate, or calcium acetate.
- Pemetrexed is a novel antifolate antimetabolite with multiple enzyme targets involved in both pyrimidine synthesis and purine synthesis and has been demonstrated to have antitumor activity on a variety of solid tumors, including mesothelioma, non-small cell lung cancer, breast, cervical, colorectal, head, neck, and bladder cancer.
- the clinical use of pemetrexed has been limited due to problems such as rapid elimination, low bioavailability, poor tumor cell selectivity, and penetration.
- liposomes the drug delivery system, to solve these problems, but liposomes also have some limitations. First, they generally show a short circulating half-life after intravenous administration. Second, they tend to stick to each other and combine to form larger vesicles in the suspension, which can lead to incubation leakage. Therefore, stability is a common problem in lipid vesicles.
- Present invention relates to a liposome which meets the aforementioned needs, eliminates the disadvantages and provides some additional advantages, is loaded with pemetrexed, anticancer drug, and is coated with natural polymer, chitosan.
- Anionic liposomes synthesized by the invention are coated with chitosan to form a cationic liposome system.
- Living cells often carry a negative charge; therefore, liposomes with positively charged surfaces are expected to interact with these cells through electrostatic interaction, facilitating the cellular uptake of the drug.
- PMX pemetrexed
- Chitosan is a natural cationic polysaccharide with biocompatibility, biodegradability, and muco-adhesiveness, and several studies have proven its potential application as an absorption enhancing agent due to the high affinity of chitosan to cell membranes. It has been found that surface modification of liposome with chitosan improves and increases the stability and muco-adhesive properties of liposome, resulting in controlled and long-term drug release.
- liposomes to biological membranes to be used as drug transport systems provide unique opportunities to transmit drug molecules to cells or subcellular compartments unlike other systems.
- the main advantage of liposomes for drug transport applications is their biocompatibility. Liposomes have had a major impact on oncology to date due to their size, biocompatibility, biodegradability, hydrophobic and hydrophilic characteristics, low toxicity, and immunogenicity properties. Liposomes can hold hydrophilic agents in their aqueous compartments and hydrophobes in a two-layer region, making them a leading drug transport platform in cancer treatment.
- the microfluidic device will be used as the liposome preparation technique, unlike the conventional lipid film layer formation technique.
- Chitosan is a biocompatible, biodegradable, and non-toxic cationic biopolymer. Viable cells usually carry a negative charge and positively charged liposome is expected to facilitate uptake of the drug into the cell. This goal cannot be achieved since the liposomes obtained in the studies using PEG are anionic. Chitosan is polycationic and achieves this goal by forming a positive layer around negatively charged liposomes.
- a liposome system consisting of natural fatty acids is created to be used in the treatment of cancer.
- Synthetic fatty acids are used when creating the liposome system in various studies. All the fatty acids to be used in the invention are natural fatty acids.
- the liposome system is formed by using microfluidic, which has many advantages according to the traditional method. The drug is released in a controlled manner in the cancerous area with this system. The stability problems of liposomes are solved by coating them with chitosan, a biopolymer abundant in nature. Characterization studies of the created system will be carried out. Afterwards, a release study will be performed at physiological pH (7,4) and acidic pH (5,5) to mimic the tumor microenvironment.
- Liposomes were synthesized using natural phospholipids in the invention. Problems encountered due to poor stability of liposomes, low resistance to gastric pH and enzymatic degradation, rapid drug leakage before reaching the target areas were solved by coating with chitosan, a natural biopolymer. Targeted treatment is provided and the healing effect of pemetrexed is not only increased, but also its side effects are reduced thanks to this system. The most important feature that distinguishes the system from other drug carrier systems is that it is a natural system that does not contain toxic materials. In addition, it is a relatively new technology compared to other methods for the production of microfluidic nano-sized liposomes to be used in liposome production.
- This system provides the opportunity to provide treatment at higher efficiency by using a lower dose of medication. This will contribute to reducing the cost of treatment and making it more accessible.
- This formulation is expected to increase the bioavailability of pemetrexed and provide an effective treatment that will be hopeful for all cancer patients.
- the present invention relates to liposomes loaded with pemetrexed, an anti-cancer drug, and coated with chitosan on their surfaces.
- the method of synthesis of liposomes of the invention generally comprises the following steps;
- An ethanolic lipid solution was prepared using phosphatidyl choline, lecithin, and cholesterol in the ratio of (6:6:1) by weight with a final concentration of 3 mg/mL in the first stage.
- PBS Phosphate Buffered Salt Solution
- Liposome was obtained in the desired size with 15 passes at 20.000 psi pressure with the microfluidic device (Microfluidizer Processor M-l 10L).
- SURFACE WITH CHITOSAN POLYMER • An ethanolic lipid solution was prepared using phosphatidyl choline, lecithin, and cholesterol in the ratio of (6:6:1) by weight with a final concentration of 3 mg/mL in the first stage.
- Liposome was obtained in the desired size with 15 passes at 20.000 psi pressure with the microfluidic device (Microfluidizer Processor M-l 10L). • The surface of the liposomes loaded with the pemetrexed drug was closed with chitosan polymer.
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- General Health & Medical Sciences (AREA)
- Public Health (AREA)
- Pharmacology & Pharmacy (AREA)
- Veterinary Medicine (AREA)
- Life Sciences & Earth Sciences (AREA)
- Animal Behavior & Ethology (AREA)
- Medicinal Chemistry (AREA)
- Dispersion Chemistry (AREA)
- Epidemiology (AREA)
- Engineering & Computer Science (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Organic Chemistry (AREA)
- Medicinal Preparation (AREA)
Abstract
The invention relates to a liposome loaded with pemetrexed, an anti-cancer drug developed for use in the treatment of cancer, and coated with natural polymer, chitosan.
Description
INVESTIGATION OF THE POTENTIAL OF PEMETREXED-LOADED LIPOSOME
SYSTEM IN CANCER TREATMENT
Technical Field of the Invention
The invention relates to a liposome loaded with pemetrexed having a field of use in the healthcare sector in general, used as an anti-cancer drug developed for use in cancer treatment in particular, and coated with natural polymer, chitosan.
State of the Art of the Invention (Prior Art)
Cancer is one of the diseases that poses a great problem for all countries in the world today. Estimates show that 30 million people will die of cancer every year by 2030. Surgery, radiation, chemotherapy, and immunotherapy are the four main methods used for cancer treatment worldwide.
Chemotherapy is one of the methods used in the treatment of cancer. However, high toxicity, poor oral bioavailability, low therapeutic indices, non-specific biodistribution, and the effect of an anticancer drug on both normal and cancerous cells are the main deficiencies of the chemotherapy method. Liposomes, which are among the drug carrier systems, are used to eliminate or minimize the negative effects occurring in traditional treatment methods, to increase reaching cellular levels, to provide effective and reliable treatment at low dosages, to eliminate or minimize toxic effects and immunogenic factors, and to obtain the desired pharmacological response in the targeted area.
Liposomes are used as a drug delivery system for different chemotherapies in the present art. Hydrophilic drugs are added to the aqueous phase during liposome synthesis, whereas lipophilic drugs are added to the organic phase. Drugs can be loaded into liposomes via active or passive loading methods. Drugs are first dissolved in water or in an organic phase; afterwards, drug-containing liposomes are prepared by appropriate methods in the passive loading method. Active drug loading means that drugs are loaded into preformed liposomes. The drug can be actively loaded into the aqueous nucleus of a liposome by means of a
chemical gradient, such as pH, ammonium sulfate, or calcium acetate. There are also liposome systems where chitosan is used as a coating material. However, there is no system in which pemetrexed-loaded liposomes are coated with chitosan. Drug leakage occurs due to the poor stability of the liposome, which limits their applicability in pemetrexed-loaded liposome systems (PLS), which are formed without coating. Coated PLS is also available. Polyethylene glycol (PEG) is used as a coating material, and as a result, PEG-coated anionic liposomes are formed.
Pemetrexed is a novel antifolate antimetabolite with multiple enzyme targets involved in both pyrimidine synthesis and purine synthesis and has been demonstrated to have antitumor activity on a variety of solid tumors, including mesothelioma, non-small cell lung cancer, breast, cervical, colorectal, head, neck, and bladder cancer. However, the clinical use of pemetrexed has been limited due to problems such as rapid elimination, low bioavailability, poor tumor cell selectivity, and penetration. There are several studies using liposomes, the drug delivery system, to solve these problems, but liposomes also have some limitations. First, they generally show a short circulating half-life after intravenous administration. Second, they tend to stick to each other and combine to form larger vesicles in the suspension, which can lead to incubation leakage. Therefore, stability is a common problem in lipid vesicles.
Brief Description and Objectives of the Invention
Present invention relates to a liposome which meets the aforementioned needs, eliminates the disadvantages and provides some additional advantages, is loaded with pemetrexed, anticancer drug, and is coated with natural polymer, chitosan.
Anionic liposomes synthesized by the invention are coated with chitosan to form a cationic liposome system. Living cells often carry a negative charge; therefore, liposomes with positively charged surfaces are expected to interact with these cells through electrostatic interaction, facilitating the cellular uptake of the drug. Compared to other chemotherapeutic drugs, non-small cell lung cancer, pemetrexed (PMX) significantly prolongs the patient's survival and is well tolerated.
The controlled release of the drug is provided by the liposome system created using natural
phospholipids, and the stability problems of the liposome are overcome by the natural polymer, chitosan. Chitosan is a natural cationic polysaccharide with biocompatibility, biodegradability, and muco-adhesiveness, and several studies have proven its potential application as an absorption enhancing agent due to the high affinity of chitosan to cell membranes. It has been found that surface modification of liposome with chitosan improves and increases the stability and muco-adhesive properties of liposome, resulting in controlled and long-term drug release.
The similarities of liposomes to biological membranes to be used as drug transport systems provide unique opportunities to transmit drug molecules to cells or subcellular compartments unlike other systems. The main advantage of liposomes for drug transport applications is their biocompatibility. Liposomes have had a major impact on oncology to date due to their size, biocompatibility, biodegradability, hydrophobic and hydrophilic characteristics, low toxicity, and immunogenicity properties. Liposomes can hold hydrophilic agents in their aqueous compartments and hydrophobes in a two-layer region, making them a leading drug transport platform in cancer treatment. The microfluidic device will be used as the liposome preparation technique, unlike the conventional lipid film layer formation technique. The production of large vesicles and then the size reduction process is applied in the traditional method. Such liposome production methods lack industrial scalability and encapsulation efficiency is generally low even though this method is widely adopted on a laboratory scale. In summary, limitations of traditional preparation methods include the complexity and length of procedures, low drug encapsulation effectiveness, and heterogeneous size distributions. Physicochemical properties of nanomaterials such as size, shape, structure, hardness, and surface modification are closely linked to the release efficiency and release profile of drugs. This device has a structure that allows a controlled mixing of lipids and organic solvent containing aqueous buffer, so that it can control these properties (size, shape, etc.). In addition, this system is suitable for using solvents (ethanol) with relatively low toxicity compared to others. Even though there are studies using microfluidic device in liposome synthesis, their number is fairly limited. The study including the liposome system created by the M-110P model microfluidics used in this invention is almost nonexistent. In addition, liposomes will be coated with chitosan, a biopolymer, and a natural system that does not harm human health will be obtained from other systems. Various studies in which pemetrexed is transported using liposomes are known in the art. There are also studies in which chitosan is used to cover the liposome surface. However, there is no system in which pemetrexed-loaded
liposomes are coated with chitosan. Stability in liposomes is a general problem. Coating with synthetic or natural polymers is required in order to prevent this problem. PEG, a synthetic polymer, was generally used for coating in PLS and anionic liposomes were obtained. Chitosan is a biocompatible, biodegradable, and non-toxic cationic biopolymer. Viable cells usually carry a negative charge and positively charged liposome is expected to facilitate uptake of the drug into the cell. This goal cannot be achieved since the liposomes obtained in the studies using PEG are anionic. Chitosan is polycationic and achieves this goal by forming a positive layer around negatively charged liposomes.
A liposome system consisting of natural fatty acids is created to be used in the treatment of cancer. Synthetic fatty acids are used when creating the liposome system in various studies. All the fatty acids to be used in the invention are natural fatty acids. In addition, the liposome system is formed by using microfluidic, which has many advantages according to the traditional method. The drug is released in a controlled manner in the cancerous area with this system. The stability problems of liposomes are solved by coating them with chitosan, a biopolymer abundant in nature. Characterization studies of the created system will be carried out. Afterwards, a release study will be performed at physiological pH (7,4) and acidic pH (5,5) to mimic the tumor microenvironment.
Liposomes were synthesized using natural phospholipids in the invention. Problems encountered due to poor stability of liposomes, low resistance to gastric pH and enzymatic degradation, rapid drug leakage before reaching the target areas were solved by coating with chitosan, a natural biopolymer. Targeted treatment is provided and the healing effect of pemetrexed is not only increased, but also its side effects are reduced thanks to this system. The most important feature that distinguishes the system from other drug carrier systems is that it is a natural system that does not contain toxic materials. In addition, it is a relatively new technology compared to other methods for the production of microfluidic nano-sized liposomes to be used in liposome production.
This system provides the opportunity to provide treatment at higher efficiency by using a lower dose of medication. This will contribute to reducing the cost of treatment and making it more accessible. This formulation is expected to increase the bioavailability of pemetrexed and provide an effective treatment that will be hopeful for all cancer patients.
Detailed Description of the Invention
The present invention relates to liposomes loaded with pemetrexed, an anti-cancer drug, and coated with chitosan on their surfaces.
The method of synthesis of liposomes of the invention generally comprises the following steps;
• Synthesis of liposomes using natural phospholipids,
• Loading the pemetrexed drug to the synthesized liposomes,
• Coating the surface of liposomes loaded with pemetrexed drug with chitosan polymer.
The process steps of the said method are described in detail below.
SYNTHESIS OF LIPOSOMES USING NATURAL PHOSPHOLIPIDS
• An ethanolic lipid solution was prepared using phosphatidyl choline, lecithin, and cholesterol in the ratio of (6:6:1) by weight with a final concentration of 3 mg/mL in the first stage.
• Vortex and bath-type sonicator were used to fully dissolve the lipids.
• Phosphate Buffered Salt Solution (PBS) was added to the ethanolic lipid solution in a ratio of 10:1 (aqueous phase:alcohol phase) and the final volume was obtained as 50 mL. It was vortexed to ensure homogeneous distribution
• Liposome was obtained in the desired size with 15 passes at 20.000 psi pressure with the microfluidic device (Microfluidizer Processor M-l 10L).
Table 1. The zeta potential and size measurement results of the empty liposome.
LOADING THE PEMETREXED DRUG TO THE LIPOSOMES AND COATING THE
SURFACE WITH CHITOSAN POLYMER
• An ethanolic lipid solution was prepared using phosphatidyl choline, lecithin, and cholesterol in the ratio of (6:6:1) by weight with a final concentration of 3 mg/mL in the first stage.
• Vortex and bath-type sonicator were used to fully dissolve the lipids • PBS 10:1 (aqueous phase: alcohol phase) and pemetrexed solution were added to the ethanolic lipid solution. Thus, a pre-emulsion containing the drug substance was prepared. It was vortexed to ensure homogeneous distribution
• Liposome was obtained in the desired size with 15 passes at 20.000 psi pressure with the microfluidic device (Microfluidizer Processor M-l 10L). • The surface of the liposomes loaded with the pemetrexed drug was closed with chitosan polymer.
Table 2. Zeta potential and size measurement results of liposomes containing different concentrations of pemetrexed
Claims
CLAIMS A liposome characterized in that it is loaded with pemetrexed, anti-cancer drug, and its surface is covered with chitosan. A method of preparing the liposome according to claim 1, characterized in that it comprises the following steps;
• Synthesis of liposomes using natural phospholipids,
• Loading the pemetrexed drug to the synthesized liposomes,
• Coating the surface of liposomes loaded with pemetrexed drug with chitosan polymer. A method according to claim 2, characterized in that the liposome is prepared with a microfluidic device with 15 passes at a pressure of 20.000 psi. A method according to claim 2, characterized in that the natural phospholipids are phosphatidyl choline, lecithin, and cholesterol. A liposome according to claim 1, characterized in that it is used as a drug carrier in the treatment of cancer.
7
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| US20170127712A1 (en) * | 2013-06-20 | 2017-05-11 | James John YIANNIOS | Dietary supplement compositions with enhanced delivery matrix, gummies, chocolates, atomizers and powders containing same, and methods of making same |
| CN106821985A (en) * | 2017-02-28 | 2017-06-13 | 福州大学 | A kind of oxygen carrying of aptamers modification carries the multi-functional liposome complex of medicine |
| WO2019157138A1 (en) * | 2018-02-07 | 2019-08-15 | L.E.A.F. Holdings Group Llc | Alpha polyglutamated pemetrexed and uses thereof |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20170127712A1 (en) * | 2013-06-20 | 2017-05-11 | James John YIANNIOS | Dietary supplement compositions with enhanced delivery matrix, gummies, chocolates, atomizers and powders containing same, and methods of making same |
| CN106821985A (en) * | 2017-02-28 | 2017-06-13 | 福州大学 | A kind of oxygen carrying of aptamers modification carries the multi-functional liposome complex of medicine |
| WO2019157138A1 (en) * | 2018-02-07 | 2019-08-15 | L.E.A.F. Holdings Group Llc | Alpha polyglutamated pemetrexed and uses thereof |
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