KR20130089213A - Solid lipid nanoparticle including elastin-like polypeptide and use thereof - Google Patents

Abstract

PURPOSE: A solid lipid nanoparticle is provided to offer an effective method for transferring an active ingredient to the target site of body. CONSTITUTION: A solid lipid nanoparticle comprises an elastin-like polypeptide in which the moiety having more than one hydrophobic groups are bound and lipid molecule. The moiety having hydrophobic group is the saturated or unsaturated hydrocarbon group, the saturated or unsaturated acyl group, the substitute amide group having a chemical formula -C(CO)N(R1)(R2), which means the R1 and R2 are the respectively independent saturated or unsaturated hydrocarbon, or the saturated or unsaturated alkoxy group. The lipid molecule is neutral lipid molecule or amphiphilic lipid molecule. The elastic-like polypeptide includes more than one repeat unit selected from among VPGXG, PGXGV, GXGVP, XGVPG, GVPGX, and a group comprised of combination thoseof more than once. The V, P, G, and X is respectively means valine, proline, glycine, and an amino acid which is not proline. [Reference numerals] (AA) Coefficient

Description

Solid lipid nanoparticles, including elastin-like polypeptides and use thereof, including elastin-like polypeptides

Elastin-like polypeptides are for solid lipid nanoparticles and their use.

Solid lipid nanoparticles (SLNs) have been developed as alternative carrier systems for emulsions, liposomes, and polymeric nanoparticles. Solid lipid nanoparticles provide benefits including film formation on the skin, including the stabilization, controlled release, occlusivity, and in vivo effects of the incorporated compounds. In addition, when using a biological lipid (biological lipid), there is an advantage that the biocompatibility and biodegradability is excellent, and the storage stability is high.

Solid lipid nanoparticles can be prepared by melting / solidification, in which case lipid molecules are melted, dispersed in water and cooled to solidify the solid lipid nanoparticles. Solid lipid nanoparticles can also be produced using emulsion fixation similar to the formation of polymeric microparticles. In this case, the lipid molecules are dissolved in a solvent, emulsified, and dispersed in an aqueous solution containing an emulsifier to cure the solid lipid nanoparticles. The role of the emulsifier may stabilize the solid lipid nanoparticles.

However, solid lipid nanoparticles are too stable to take a long time for drug release. In order to overcome this, there is a need for a method that can effectively release the drug.

One aspect is to provide a solid lipid nanoparticle comprising an elastin-like polypeptide.

Another aspect is to provide a pharmaceutical composition comprising solid lipid nanoparticles comprising an elastin-like polypeptide comprising the active ingredient.

Another aspect is to provide a method for efficiently delivering an active ingredient to a target site in a subject's body using the solid lipid nanoparticles.

One aspect provides a solid lipid nanoparticle comprising an elastin-like polypetide (ELP) and a lipid molecule conjugated with a moiety comprising a hydrophobic group.

The term "elastin-like polypeptide" refers to a class of amino acid polymers that undergo conformational changes with temperature. In one embodiment, the elastin-like polypeptide can be a polymer having "inverse phase transitioning behavior". Reverse phase transition behavior indicates that the solution is soluble in aqueous solutions at temperatures below the "inverse phase transition temperature (T _t )" and becomes insoluble when the temperature rises above the reverse phase transition temperature. Elastin-like polypeptides can transition from highly soluble elongated chains to tightly folded aggregates with greatly reduced solubility as the temperature rises. This reverse phase shift may be induced by having more β-turns and a distorted β-structure as the structure of the elastin-like polypeptide increases with increasing temperature. The elastin-like polypeptide may be, for example, having a phase transition temperature of about 10 ° C to about 70 ° C or about 39 ° C to about 70 ° C.

In the solid lipid nanoparticles, the reverse phase transition behavior destroys the solid lipid nanoparticles due to shrinkage and self-assembly of the elastin-like polypeptide as the temperature rises from a temperature lower than the T _t of the elastin-like polypeptide. can do. Destruction of the solid lipid nanoparticles may increase the release of the active material contained in the solid lipid nanoparticles. Thus, the active ingredient included in the solid lipid nanoparticles can be released from the solid lipid nanoparticles to the outside with higher sensitivity to temperature. However, one or more embodiments are not limited to specific mechanisms of action.

The destruction of the solid lipid nanoparticles by the reverse phase transition behavior of the elastin-like polypeptide may depend on the phase transition temperature of the elastin-like polypeptide and the lipid molecules constituting the solid lipid nanoparticles. Lipid molecules are present in the gel phase below their phase transition temperature and in the liquid (crystalline) phase at higher temperatures. When the lipid molecules are present in the gel, even if the elastin-like polypeptide is changed to a structure having a β-turn structure by reverse phase transition behavior, destruction of the solid lipid nanoparticles may not occur or may be limited. On the other hand, when lipid molecules are present in the liquid phase, destruction of the solid lipid nanoparticles can be induced as the elastin-like polypeptide is changed to a structure having a β-turn structure by reverse phase transition behavior. That is, the reverse phase transition of elastin-like polypeptides induces the destruction of solid lipid nanoparticles more efficiently when the lipid molecules are in the liquid phase as compared to the gel phase. Thus, the release temperature of the active ingredient included in the solid lipid nanoparticles can be controlled by controlling the phase transition temperature of the lipid molecules constituting it and the reverse phase transition temperature of the elastin-like polypeptide. For example, the phase transition temperatures of the elastin-like polypeptide, the lipid molecules constituting the solid lipid nanoparticles, and the solid lipid nanoparticles are about 10 ° C. to 70 ° C., for example, 35 ° C. to 70 ° C., 39 ° C. to 45 ° C. Or 39 ° C. to 60 ° C.

Elastin-like polypeptides have a moiety comprising a hydrophobic group at the N-terminal, C-terminal, N-terminal amino acid residue side chains of the amino acid sequence, the side chains of the C-terminal amino acid residues, or the side chains of amino acid residues which are not between the N and C termini. It may be connected to. For example, the moiety including the hydrophobic group may be conjugated to the N terminal or the C terminal, and one or more molecules per one elastin-like polypeptide molecule are conjugated. For example, a moiety comprising a hydrocarbon group and the hydrophobic group such as a carbonyl-containing group such as an acyl group and an alkoxy group may be conjugated through an amine or amide bond with an N-terminal N atom or an ester bond with a C-terminal carbonyl group. It may have been. Here, the moiety including the hydrophobic group may be a carbonyl-containing group such as C4-C30, for example, a hydrocarbon having an carbon number of C14-C24, or a carbon number of C16-C24, or an acyl group and an alkoxy group.

In one embodiment, the elastin-like polypeptide can be defined by an amino acid sequence. The elastin-like polypeptide is partially or entirely composed of VPGXG (SEQ ID NO: 1), PGXGV (SEQ ID NO: 2), GXGVP (SEQ ID NO: 3), XGVPG (SEQ ID NO: 4), GVPGX (SEQ ID NO: 5), and combinations thereof At least one repeating unit selected from the group, wherein V is valine, P is proline, G is glycine, and X is a natural or non-natural amino acid that is not proline. Here, X of each repeating unit may be the same or different amino acids. The repeating units may be separated from each other by one or more amino acids that do not remove the phase transfer properties of the resulting elastin-like polypeptide, or the terminal portion of the ELP may comprise one or more amino acids, for example amino acids other than the repeating units. The molar ratio of the repeating unit to another amino acid may be about 0.1% to about 99.9% of the repeating unit based on the amino acid different from the repeating unit. The selected repeating unit may be repeated two or more times, for example, 2 to 200 times.

The elastin-like polypeptide is a tandemly-repeated block of one or more series of VPGXG, PGXGV, GXGVP, XGVPG, GVPGX, or a combination thereof, or VPGXG, PGXGV, GXGVP, XGVPG, GVPGX or a combination thereof. It may include a block repeated in series more than once. The elastin-like polypeptide is composed not only of VPGXG, PGXGV, GXGVP, XGVPG, GVPGX, or a combination thereof, as long as reverse phase transition behavior is maintained, as well as alanine at the terminal end and / or both ends of the repeating unit. It may include one or more amino acids such as glycine. The elastin-like polypeptide may be one in which the N- or C-terminal is bonded to a moiety including a hydrophobic group. In addition, moieties comprising hydrophobic groups can be conjugated by binding to reactive groups in the side chain of the amino acid residue of the elastin-like polypeptide. The reactive group may be an amino group, hydroxyl group, thiol group or carboxyl group. Opposite ends that are not bound to a moiety comprising a hydrophobic group may or may not be blocked. For example, when a moiety comprising a hydrophobic group and an elastin-like polypeptide are bound through the N terminus of an elastin-like polypeptide, the carboxyl group at the C terminus of the elastin-like polypeptide may be blocked or unblocked. The blocking may be by binding to or reacting with a material that is biocompatible, non-immunogenic, aids in specific delivery, or may be avoided from a biodegradation system. For example, the blocking can be accomplished by forming an amide bond by bonding an amino group with a carboxyl group at the C terminus of an elastin-like polypeptide. The amino group may be derived from an ammonia molecule, primary amine, secondary amine or tertiary amine. The primary amine, secondary amine or tertiary amine may be each having a carbon number of C1-C18, for example C1-C6. X may be valine or alanine.

The repeating unit may be included by repeating one or more integer times independently of each other. The repetition frequency is independently 2 to 200, 2 to 100, 2 to 80, 2 to 60, 2 to 40, 2 to 10, 2 to 12, 2 to 8, 2 to 6, 4 to 100, 8 to 80 , 10 to 60, 12 to 40, 20 to 40, 4 to 10, 4 to 8, or may be an integer of 4 to 6.

The moiety comprising the hydrophobic group may be a molecule having a property of interacting with the lipid molecules to fix an elastin-like polypeptide to the lipid molecule. The interaction may be a covalent or noncovalent bond. The interaction may be hydrophobic interaction, van der Waals bond, ionic bond, or hydrogen bond. The moiety including the hydrophobic group may be one or all of the same or different lipid molecules with the lipid molecules.

The moiety including the hydrophobic group may include a molecule composed only of a hydrophobic portion or an amphiphilic molecule including a hydrophobic portion and a hydrophilic portion. The amphiphilic molecule may be a hydrophobic portion is disposed in the lipid molecule and the hydrophilic portion is disposed in the outer direction of the lipid molecule is bound to the elastin-like polypeptide. Here, the "outer direction" refers to the outward direction from the center of the solid lipid nanoparticles. The moiety may be packed, embedded or dispersed between the lipid molecules in the solid lipid nanoparticles. The moiety may be a lipid molecule naturally present in the biological membrane or a lipid molecule although not naturally present in the biological membrane.

Lipid molecules naturally present in the biological membrane may be selected from the group consisting of phospholipids or derivatives thereof, sterols or derivatives thereof, sphingolipids or derivatives thereof, and combinations thereof. The phospholipid or derivative thereof may be one or more selected from the group consisting of phosphatidyl choline, phosphatidyl glycerol, phosphatidyl inositol, phosphatidyl ethanolamine, and combinations thereof. The sterol or derivatives thereof may be cholesterol or derivatives thereof, or squalene or derivatives thereof. The sphingolipids may be sphingomyelin or gangliosides or derivatives thereof. The moiety includes an intermediate or precursor produced during in vivo synthesis of the phospholipid, sterol or sphingolipid. For example, phosphoglycerides, spingosin, ceramides or cerebrosides. The derivative may be an ester of a fatty acid. The fatty acid may be a C4 to C30 fatty acid.

The moiety is a saturated or unsaturated hydrocarbon group, a substituted amide group of the formula -C (CO) N (R1) (R2), saturated or unsaturated acyl group or saturated or unsaturated, wherein R1 and R2 are each independently saturated or unsaturated hydrocarbons. It may be an alkoxy group.

Conjugation of an elastin-like polypeptide with a moiety comprising a hydrophobic group may be one that is not cleavable under physiological or pathological conditions, or is made by cleavable linkage. Examples of cleavable linkages may be linkages made through a pH cleavable linker, a heat transferable linker, a radiation cleavable linker, a linker delivered in an aqueous solution.

The moiety may be conjugated by binding to an NH or C-terminal carbonyl (-C (O)-) group at the N-terminal elastin-like polypeptide. The moiety may be linked by a bond formed by reaction with a functional group selected from the group consisting of side groups of the elastin-like polypeptide, for example, an amino group, a carbonyl group, a hydroxy group, a thiol group, and a combination thereof. . The moiety may be conjugated via an amine or amide bond with the N atom of the elastin-like polypeptide. The moiety including the hydrophobic group may be conjugated to the carbonyl group of the C terminal of the elastin-like polypeptide through an amide or ester bond.

The moiety may be a hydrophobic group moiety having C4-C30, for example, C14-C24 carbon number, or C16-C24 carbon number. The moiety can be, for example, myristoyl (C14), palmitoyl (C16), stearoyl (C18), arachidonyl (C20), behenonyl (C22) or lignoseroyl (C24); Or myristyl (C14), palmityl (C16), stearyl (C18), arachidyl (C20), behenyl (C22) or lignoceryl (C24). R1 and R2 of the substituted amide group of formula -C (CO) N (R1) (R2) are myristyl (C14), palmityl (C16), stearyl (C18), arachidyl (C20), behenyl ( C22) or lignoceryl (C24). The moiety can be filled with lipid molecules by a hydrophobic effect, such that an elastin-like polypeptide conjugated to the moiety can be immobilized on the solid lipid nanoparticles.

An example of an ELP conjugated with a moiety comprising a hydrophobic group may be stearoyl- or cholesteryl-V′n-NH 2 with n of 1 to 200. Here, V ′ represents any one of VPGXG, PGXGV, GXGVP, XGVPG, and GVPGX, and when n is 2 or more, V ′ included in the sequence may be the same or different sequences. n is 2 to 200, 2 to 100, 2 to 80, 2 to 60, 2 to 40, 2 to 10, 2 to 12, 2 to 8, 2 to 6, 4 to 100, 8 to 80, 10 to 60, It may be an integer of 12 to 40, 20 to 40, 4 to 10, 4 to 8, or 4 to 6. Here, V may be valine, P is proline, G is glycine, and X may be a natural or non-natural amino acid other than proline. Here, X of each repeating unit may be the same or different amino acids. The ELP conjugated with the moiety comprising a hydrophobic group can be, for example, stearoyl- (VPGVG) n-NH 2 or cholesteryl- (VPGVG) n-NH 2.

Another example of an ELP conjugated with a moiety comprising a hydrophobic group is that sterolyl- or cholesteryl- [V ₁ n ₁ V ₂ n ₂ ] n ₃ where n ₁ , n ₂ and n ₃ are each 1 to 200; -NH ₂ may be. 2 to 200, 2 to 100, 2 to 80, 2 to 60, 2 to 40, 2 to 10, 2 to 12, 2 to 8, 2 to 6, 4 to 100, 8 to 80, 10 to 60, 12 to 40, 20 to 40, 4 to 10, 4 to 8, or may be an integer of 4 to 6. Where V ₁ And V ₂ represents any one of VPGXG, PGXGV, GXGVP, XGVPG, and GVPGX, respectively. Here, V may be valine, P is proline, G is glycine, and X may be a natural or non-natural amino acid other than proline. Here, X of each repeating unit may be the same or different amino acids. The ELP conjugated with the moiety comprising a hydrophobic group can be, for example, stearoyl- or cholesteryl-[(VPGVG) n ₁ (VPGAG) n ₂ ] n ₃ -NH ₂ .

Other examples of ELP conjugated moieties comprising hydrophobic groups include stearyl- or cholesteryl- [B (SA or Chol) n ₁ V ₁ n wherein n ₁ , n ₂ and n ₃ are each 1 to 200; ₂ ] n ₃ -NH ₂ . n ₁ , n ₂ and n ₃ are 2 to 200, 2 to 100, 2 to 80, 2 to 60, 2 to 40, 2 to 10, 2 to 12, 2 to 8, 2 to 6, 4 to 100, It may be an integer of 8 to 80, 10 to 60, 12 to 40, 20 to 40, 4 to 10, 4 to 8, or 4 to 6. Wherein B (SA or Chol) is any one of VPGXG, PGXGV, GXGVP, XGVPG, and GVPGX, wherein V is valine, P is proline, G is glycine, and X is stearoyl- or cholesteryl-side chain Lysine, arginine or histidine with an amino group. V ₁ is any one of VPGXG, PGXGV, GXGVP, XGVPG, and GVPGX, where V is valine, P is proline, G is glycine, and X is a natural or non-natural amino acid that is not proline. Here, when n1 or n2 is two or more numbers, B of each position may be the same or different, and V ₁ of each position may be the same or different, respectively. The ELP conjugated with the hydrophobic group may be, for example, stearoyl- or cholesteryl-[(VPGK (SA or Chol) G) n ₁ (VPGXG) n ₂ ] n ₃ -NH ₂ . .

The elastin-like polypeptide may be included in a ratio of 0.01 to 50% by weight relative to the total weight of the solid lipid nanoparticles.

As used herein, the term “lipid molecule” includes fats or fat-derived materials that are relatively insoluble in water but are soluble in organic solvents. Lipid molecules include fatty acid esters, fatty alcohols, sterols, waxes and the like. Examples of such fats are glyceryl esters of higher fatty acids.

The lipid molecules include neutral lipid molecules or amphipathic lipid molecules. Neutral lipid molecules represent lipid molecules with no charge. Neutral lipid molecules may be glyceryl esters of fatty acids. Neutral lipid molecules include monoglycerides, diglycerides or triglycerides of one or more C4 to C24 carboxylic acids. The carboxylic acid may be saturated or unsaturated and branched or unbranched. For example, the lipid molecules are C4, C5, C6, C7, C8, C9, C10, C11, C12, C13, C14, C15, C16, C17, C18, C19, C20, C21, C22, C23, C24 Monoglycerides of acids. The carboxylic acid may be saturated or unsaturated and branched or unbranched. The carboxylic acid may be covalently linked to any of the three glycerol hydroxyl groups. In another embodiment, the lipid molecule is C4, C5, C6, C7, C8, C9, C10, C11, C12, C13, C14, C15, C16, C17, C18, C19, C20, C21, C22, C23, C24 Diglycerides of carboxylic acids. The carboxylic acid may be saturated or unsaturated and branched or unbranched. The two carboxylic acids may be the same or different and may be covalently linked to any two of the three glycerol hydroxyl groups. In another embodiment, the lipid molecule is C4, C5, C6, C7, C8, C9, C10, C11, C12, C13, C14, C15, C16, C17, C18, C19, C20, C21, C22, C23, C24 Triglycerides of carboxylic acids. The carboxylic acid may be saturated or unsaturated and branched or unbranched. The three carboxylic acids may be the same, the two carboxylic acids may be the same, or all three may be different. It may be covalently linked to any two of the three glycerol hydroxyl groups.

In one embodiment, the lipid molecule may be a blend of triglycerides of saturated even, unbranched fatty acids having a chain length of C8 to C18. For example, the lipid molecule may be a mixture of each triglyceride of C8, C10, C12, C14, C16, or C18 carboxylic acid. For example, it may be a mixture of tricapric glycerol and trilauric glycerol. For each triglyceride in the mixture, each carboxylic acid may be the same, two of the carboxylic acids may be the same, or all three may be different.

The lipid molecule may comprise a mixture of monoglycerides, diglycerides and triglycerides. Each carboxylic acid of monoglycerides, diglycerides and triglycerides may be saturated or unsaturated, branched or unbranched, C4, C5, C6, C7, C8, C9, C10, C11, C12, C13, C14, C15, C16 , C17, C18, C19, C20, C21, C22, C23, C24 carboxylic acid. The lipid molecule may be a mixture of monoglycerides, diglycerides and triglycerides of saturated even, unbranched fatty acids having a chain length of C8 to C18. For example, the lipid molecule may be a mixture of each triglyceride of C8, C10, C12, C14, C16, or C18 carboxylic acid.

Amphiphilic lipid molecules can include hydrophilic and hydrophobic moieties. Amphiphilic lipid molecules can be molecules with a hydrophilic head and a hydrophobic tail. Amphiphilic lipid molecules may comprise one or more selected from the group consisting of phospholipids, fatty acids and combinations thereof. The lipid molecule may be one having a carbon atom of C14 to C50. The lipid molecule may be a phospholipid. The phospholipid may be one having a carbon atom of C12 to C24. The phospholipid may be one or more selected from the group consisting of phosphatidyl choline, phosphatidyl glycerol, phosphatidyl inositol, and phosphatidyl ethanolamine. The at least one phospholipid may have one acyl group. In addition, the phospholipid may be one having a phase transition temperature of about 10 ℃ to about 70 ℃, for example, about 39 ℃ to about 60 ℃ or about 38 ℃ to about 45 ℃. The phospholipid may be saturated or unsaturated acyl group. The phospholipid may be a mixture of two or more phospholipid molecules. Mixing two or more phospholipid molecules can produce solid lipid nanoparticles having various phase transition temperatures.

Phospholipid molecules include, for example, two acyl groups, C12 saturated chain phospholipids (Tc = about 10 ° C.), C14 saturated chain phospholipids (Tc = about 24 ° C.), C16 saturated chain phospholipids (Tc = about 41 ° C), C18 saturated chain phospholipids (Tc = about 55 ° C), C20 saturated chain phospholipids (Tc = about 65 ° C), C22 saturated chain phospholipids (Tc = about 70 ° C), and combinations thereof It may be one or more selected. Similar to the phase transition temperatures at which phosphatidylcholine changes in a similar fashion along the length of their acyl chains, phosphatidylglycerol, phosphatidyl inositol, phosphatidyl ethanolamine, spingomyelin and gangliosides can also be used. Phosphatidylcholine can be egg phosphatidylcholine.

An example of a C16 saturated chain phospholipid may be dipalmitoylphosphatidylcholine (DPPC). DPPC is a saturated chain (C16) phospholipid with a phase transition temperature of about 41.5 ° C. An example of a C18 saturated chain phospholipid may be 1,2-distearoyl-sn-glycero-3-phosphocholine (1,2-distearoyl-sn-glycero-3-phosphocholine (DSPC)). DSPC is a saturated chain (C18) phospholipid with a phase transition temperature of about 55.10 ° C.

The lipid molecule may include other membrane-forming substances in addition to phospholipids. Exemplary materials that form solid membranes can be boll lipids or bacterial lipids. In addition, block copolymers comprising water soluble polymers (eg polyethylene glycol) and water insoluble polymers (eg polypropylene oxide and polyethylethylene) can be used. In addition, the lipid molecules include polymeric lipid molecules. For example, the lipid molecule may be esterified poly (acrylic acid) or esterified poly (vinyl alcohol).

The lipid molecule or a mixture of lipid molecules is 0.01 to 90 (0.5, 1, 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, based on the total weight of the solid lipid nanoparticles) 65, 70, 75, 80, or 85) weight percent, for example, 0.5-90 wt%, 1.0-90 wt%, 5.0-90 wt%, 10.0-90 wt%, 15.0-90 wt%, 30.0-90 wt%, 50.0- 90 wt%, 75.0-90 wt%, 80.0-90 wt%, or 10.0-80 wt%.

The term "solid" indicates that at least a portion of the solid lipid nanoparticles are solid at room temperature and atmospheric pressure. However, solid lipid nanoparticles may comprise liquid lipids and / or entrained solvents.

The solid lipid nanoparticles may further comprise stabilizers thereof. The stabilizer may be selected from the group consisting of sterols or derivatives thereof, sphingolipids or derivatives thereof and combinations thereof. The stabilizer is cholesterol, β-cholesterol, fatty acid esters of cholesterol, for example fatty acid esters of C10 to C24, cystosterol, ergosterol, stigmasterol, 4,22-stigmasteradien-3-one, stigmasterol Acetate, lanosterol, and combinations thereof.

The solid lipid nanoparticle stabilizer may be selected from the group consisting of steroids, sphingolipids or derivatives thereof and combinations thereof. The stabilizer may be a steroid having a property that can be incorporated (incorporation) between lipid molecules. As used herein, the term "steroid" refers to a form of four fused cycloalkane rings, namely three cyclohexane rings and one cyclopentane ring, designated A, B and C rings from left to right (D Gonane or an organic compound comprising a skeleton derived therefrom. As used herein, the "skeleton derived therefrom" includes the introduction of an unsaturated bond into the scaffold. The steroid may vary depending on the functional groups attached to the four rings and the oxidation state of the ring. For example, the steroid may be to include a hydrophilic functional group in the ring. For example, the steroid may be one having a hydroxyl group in the ring. The steroid may be sterol. The term "sterol" refers to a form of steroids having a skeleton derived from cholestane having a hydroxyl group at position 3. As used herein, the term "derived backbone" includes the introduction of unsaturated bonds in the cholestan backbone. Such steroids include steroids that are found in plants, animals and fungi. For example, it may be made from cycloartenol as in lanosterol or in plants as in animals or fungi. The sterols include cholesterol or derivatives thereof. As used herein, "derivative" means a derivative of cholesterol that retains its property of being incorporated into a lipid bilayer. The derivatives include fatty acid esters. The stabilizer may be selected from the group consisting of cholesterol, cystosterol, ergosterol, stigmasterol, 4,22-stigmasteradien-3-one, stigmasterol acetate, lanosterol, cycloarthenol, and combinations thereof. have.

The stabilizer is from 0 to 50% by weight, for example 1 to 50% by weight, 5 to 50% by weight, 10 to 50% by weight, 20 to 50% by weight, 30 to 50% by weight relative to the total weight of the solid lipid nanoparticles %, 1 to 40% by weight, 5 to 20% by weight, 10 to 40% by weight, 20 to 30% by weight, or 1 to 10% by weight.

The solid lipid nanoparticle may further include a phospholipid derivative derivatized with a hydrophilic polymer. The hydrophilic polymer may be selected from the group consisting of polyethylene glycol, polylactic acid, polyglycolic acid, polylactic acid and polyglycolic acid copolymers, polyvinyl alcohol, polyvinylpyrrolidone, oligosaccharides, and combinations thereof. The derivative may be a PEG conjugated to a phospholipid of C4-C30, for example C16-C24. The derivative may be DPPC-PEG, or DSPE-PEG. The PEG may have a molecular weight of 180 to 50,000 Da. The derivative is 0 to 10% by weight, for example, 1 to 10% by weight, 2 to 10% by weight, 3 to 10% by weight, 5 to 10% by weight, 1 to 8% by weight based on the total weight of the solid lipid nanoparticles %, 2 to 5% by weight, 3 to 5% by weight, or 1 to 5% by weight.

Lipid molecules constituting the solid lipid nanoparticles may have a phase transition temperature of 39 ℃ to 60 ℃. The elastin-like polypeptide may be one having a phase transition temperature of 35 ℃ to 70 ℃ or 39 ℃ to 70 ℃.

The solid lipid nanoparticles may have an average diameter of about 10 nm to 1500 nm, for example, about 10 nm to 1000 nm, about 10 nm to 500 nm, about 10 nm to 300 nm, about 100 nm to 300 nm, or about 100 nm to 200 nm.

The solid lipid nanoparticles may further comprise an active substance. The active substance may further include one or more selected from the group consisting of physiologically active agents, pharmaceutically active agents, magnetically active agents, imaging agents, and combinations thereof. The pharmaceutically active ingredient is selected from the group consisting of anesthetics, antihistamines, anticancer agents, antiulcers, antiseizure agents, muscle relaxants, immunosuppressants, anti-infective agents, nonsteroidal anti-inflammatory agents, imaging agents, nutrients, and combinations thereof. Can be. The active ingredient is methotrexate, doxorubicin, epirubicin, daunorubicin, vincristine, vinblastine, etoposide, elliptisin, camptothecin, paclitaxel, docetaxel, cisplatin, prednisone, methyl-prednisone, ibuprofen and combinations thereof It may be selected from the group consisting of. The active substance may be dispersed, embedded or incorporated into solid lipid nanoparticles. The active substance is 0.01 to 10% by weight, for example, 0.1 to 10% by weight, 1 to 10% by weight, 3 to 10% by weight, 5 to 10% by weight, 0.01 to 8% by weight, based on the total weight of the solid lipid nanoparticles. %, 0.01 to 5% by weight, or 0.1 to 5% by weight.

The solid lipid nanoparticles may further comprise a surfactant. Surfactant is a substance that dissolves in liquid and significantly lowers the surface tension. It is divided into a hydrophilic part and a hydrophobic part in a molecule. Therefore, the surfactant is easily adsorbed to the interface, and is a molecule called micelle at a certain concentration (critical micelle concentration). To form an aggregate. In this system, surfactants can be used for entanglement and uniform dispersion of lipid particles. The surfactant may be a polyoxyethylene derivative (Tween) of polyethers such as sorbitan monolaurate. The surfactant can be Tween 20 or Tween 80. The surfactant is 0 to 10% by volume, for example 0 to 10% by volume, 1 to 10% by volume, 3 to 10% by volume, 5 to 10% by volume, 1 to 8% by volume relative to the total volume of solid lipid nanoparticles. %, 1 to 5% by volume, or 3 to 5% by volume. One or more surfactants may be dissolved or suspended in the organic or aqueous phase during the preparation of the solid lipid nanoparticles. In other embodiments, one or more surfactants may be added to the suspension of solid lipid nanoparticles after preparation.

If the surfactant is present in the organic and / or aqueous phase, the surfactant is internal to the solid lipid nanoparticles (ie, encapsulated or external to the solid lipid nanoparticles (ie, functional groups present on the outside). Covalently or non-covalently bound to) When one or more surfactants are mixed after the formation of the solid lipid nanoparticles, the surfactants are covalently or non-covalently bound to functional groups present outside, ie, external to, the solid lipid nanoparticles. Can be located.

One embodiment of the solid lipid nanoparticles includes an elastin-like polypeptide conjugated with a moiety including a hydrophobic group, a first lipid molecule, a second lipid molecule, and a stabilizer, wherein the first lipid molecule is a phospholipid, The second lipid molecule may be a solid lipid nanoparticle that is a neutral lipid molecule. The moieties, stabilizers, phosphorus lipid molecules and neutral lipid molecules are as described above. In the above embodiment, the phospholipid may be one or more selected from the group consisting of phosphatidyl choline, phosphatidyl glycerol, phosphatidyl inositol, phosphatidyl ethanolamine, and combinations thereof. The phospholipid may be one having a C16-C24 acyl group. The neutral lipid molecule may include one or more selected from the group consisting of at least one C4 to C24 carboxylic acid monoglyceride, diglyceride, triglyceride, and combinations thereof. The stabilizer may be selected from the group consisting of sterols or derivatives thereof, sphingolipids or derivatives thereof and combinations thereof. The stabilizers include cholesterol, β-cholesterol, C10 to C24 fatty acid esters of cholesterol, cystosterol, ergosterol, stigmasterol, 4,22-stigmagstadien-3-one, stigmasterol acetate, lanosterol and their It may be selected from the group consisting of a combination.

In one embodiment, the solid lipid nanoparticles may comprise ELP, phosphatidylcholine, triglyceride and cholesteryl oleate composed of tricaprine and trilaurin conjugated with a moiety comprising a hydrophobic group. Phosphatidylcholine, triglyceride and cholesteryl oleate consisting of tricaprine and trilaurin is a molar ratio of 2-5: 0.1-3: 0-1, and a molar ratio of tricaprine and trilaurin is from 1: 0.25 to about 4 and the moiety comprising a hydrophobic group conjugated ELP may be about 0.01 to about 50 weight percent of phosphatidylcholine. Here, an example of the ELP to which the moiety including the hydrophobic group is conjugated may be stearyl- or cholesteryl-V'n-NH ₂ where n is 1 to 200. Here, V ′ represents any one of VPGXG, PGXGV, GXGVP, XGVPG, and GVPGX, and when n is 2 or more, V ′ at each position of the sequence may be the same or different sequence. n is 2 to 200, 2 to 100, 2 to 80, 2 to 60, 2 to 40, 2 to 10, 2 to 12, 2 to 8, 2 to 6, 4 to 100, 8 to 80, 10 to 60, It may be an integer of 12 to 40, 20 to 40, 4 to 10, 4 to 8, or 4 to 6. Here, V may be valine, P is proline, G is glycine, and X may be a natural or non-natural amino acid other than proline. Here, X of V 'at each position may be the same or different amino acid. The ELP conjugated with the moiety comprising a hydrophobic group can be, for example, stearoyl- (VPGVG) n-NH ₂ or cholesteryl- (VPGVG) n-NH ₂ .

Another example of an ELP conjugated with a moiety comprising a hydrophobic group is that sterolyl- or cholesteryl- [V ₁ n ₁ V ₂ n ₂ ] n ₃ where n ₁ , n ₂ and n ₃ are each 1 to 200; -NH ₂ may be. n ₁ , n ₂ and n ₃ are 2 to 200, 2 to 100, 2 to 80, 2 to 60, 2 to 40, 2 to 10, 2 to 12, 2 to 8, 2 to 6, 4 to 100, 8 It may be an integer from to 80, 10 to 60, 12 to 40, 20 to 40, 4 to 10, 4 to 8, or 4 to 6. When n ₁ and n ₂ are each 2 or more, V ′ at each position of the sequence may be the same or different sequence. Where V ₁ And V ₂ represents any one of VPGXG, PGXGV, GXGVP, XGVPG, and GVPGX, respectively. Here, V may be valine, P is proline, G is glycine, and X may be a natural or non-natural amino acid other than proline. Here, X of V 'at each position may be the same or different amino acid. The ELP conjugated with the moiety comprising a hydrophobic group can be, for example, stearoyl- or cholesteryl-[(VPGVG) n ₁ (VPGAG) n ₂ ] n ₃ -NH ₂ .

Other examples of ELP conjugated moieties comprising hydrophobic groups include stearyl- or cholesteryl- [B (SA or Chol) n ₁ V ₁ n wherein n ₁ , n ₂ and n ₃ are each 1 to 200; ₂ ] n ₃ -NH ₂ . n ₁ , n ₂ and n ₃ are 2 to 200, 2 to 100, 2 to 80, 2 to 60, 2 to 40, 2 to 10, 2 to 12, 2 to 8, 2 to 6, 4 to 100, It may be an integer of 8 to 80, 10 to 60, 12 to 40, 20 to 40, 4 to 10, 4 to 8, or 4 to 6. Wherein B (SA or Chol) is any one of VPGXG, PGXGV, GXGVP, XGVPG, and GVPGX, wherein V is valine, P is proline, G is glycine, X is stearoyl- or cholesteryl-conjugated Lysine, arginine or histidine with side chain amino groups. V ₁ is any one of VPGXG, PGXGV, GXGVP, XGVPG, and GVPGX, where V is valine, P is proline, G is glycine, and X is a natural or non-natural amino acid that is not proline. Here, when n ₁ or n ₂ is a number of two or more, B and V ₁ at each position may be the same or different. The ELP conjugated with the hydrophobic group may be, for example, stearoyl- or cholesteryl-[(VPGK (SA or Chol) G) n ₁ (VPGXG) n ₂ ] n ₃ -NH ₂ . .

Methods of making solid lipid nanoparticles are known. Solid lipid nanoparticles can be prepared by melting / solidification, in which case lipid molecules are melted, dispersed in water and cooled to solidify the solid lipid nanoparticles. Solid lipid nanoparticles can also be produced using emulsion fixation similar to the formation of polymeric microparticles. In this case, the lipid molecules are dissolved in a solvent, emulsified, and dispersed in an aqueous solution containing an emulsifier to cure the solid lipid nanoparticles. The role of the emulsifier may stabilize the solid lipid nanoparticles. Alternatively, it may be prepared by film formation / hydration. In this case, after dissolving / removing the lipid molecule and the hydrophobic drug in an organic solvent at the same time, the solid lipid nanoparticles into which the drug is introduced may be prepared through sonication and vortexing. Depending on the composition of the solid lipid nanoparticles, two production methods may be used differently.

Solid lipid nanoparticles include, for example, providing an organic phase comprising an elastin-like polypeptide, a lipid molecule, and a binary solvent system; Removing the organic phase to form a thin film; Providing an aqueous phase comprising water; And combining and dispersing the thin film and the aqueous phase. The organic phase, or a portion of the organic phase, can be selectively removed so that the solid lipid nanoparticles can be prepared as an aqueous suspension. The term “two phase solvent system” refers to a solvent system comprising two or more miscible or partially miscible solvents. This term may especially include three solvents, four solvents, and five solvent systems. The solvent system generally includes a solvent that is liquid at room temperature and atmospheric pressure. However, while the overall system is a liquid at room temperature and atmospheric pressure, it should be understood that one or more solvents of the system may be solid or gas at room temperature and atmospheric pressure. The biphasic solvent system is chloroform (CHCl ₃ ) -ethanol, chloroform (CHCl ₃ ) -methanol, dichloromethane (CH ₂ Cl ₂ ) -ethanol, dichloromethane (CH ₂ Cl ₂ ) -methanol, N-methylpyrrolidone (NMP) -acetone, tetrahydrofuran (THF) -acetone, or dimethylformamide (DMF) -acetone.

Solid lipid nanoparticles may also comprise providing an organic phase comprising an elastin-like polypeptide, neutral lipid molecules and a binary solvent system; Removing the organic phase to form a thin film; Providing an aqueous phase comprising water; And combining and dispersing the thin film and the aqueous phase. However, the production of solid lipid nanoparticles can be any method known and is not limited to any particular method.

In one embodiment, the solid lipid nanoparticles include an elastin-like polypeptide conjugated with a moiety comprising a hydrophobic group, a first lipid molecule, a second lipid molecule, and a stabilizer, wherein the first lipid molecule is a phospholipid, The second lipid molecule may be a solid lipid nanoparticle which is a neutral lipid molecule, the second lipid molecule and the stabilizer form an inner core, and the first lipid molecule may have a structure surrounding the inner core. The first lipid molecule may surround the inner core in the form of a single layer. The elastin-like polypeptide may be immobilized to the solid lipid nanoparticle by interacting with a first lipid molecule and optionally a second lipid molecule through the moiety. The elastin-like polypeptide, the first lipid molecule, the second lipid molecule and the stabilizer are as described above. Solid lipid nanoparticles may also include surfactants.

Another aspect is a pharmaceutical composition for delivering to an individual an active ingredient comprising a pharmaceutically acceptable carrier or diluent and a solid lipid nanoparticle comprising the active ingredient, wherein the solid lipid nanoparticle comprises a hydrophobic group. It provides a composition comprising; conjugated elastin-like polypeptide and lipid molecules.

The pharmaceutically acceptable carrier or diluent may be known in the art. The carrier or diluent may be selected from the group consisting of water, for example saline and sterile water, Ringer's solution, buffers, dextrose solution, maltodextrin solution, glycerol, ethanol and combinations thereof.

The solid lipid nanoparticles are as described above.

The solid lipid nanoparticles may be dispersed in an aqueous medium. The aqueous medium may be one containing saline or PBS. In addition, the solid lipid nanoparticles can be entrapped in liposomes, or formulated in the form of dispersions or emulsions. The active ingredient may be collected in the internal space of the solid lipid nanoparticles. The active ingredient may be collected in the lipid molecules of the solid lipid nanoparticles. The solid lipid nanoparticles may have a phase transition temperature of 39 ℃ to 45 ℃.

The active ingredient may be one or more selected from the group consisting of physiologically active agents, pharmaceutically active agents, magnetically active agents, imaging agents, and combinations thereof. The active ingredient may be at least one of a water insoluble ingredient, a water soluble ingredient, and a combination thereof. The pharmaceutically active ingredient is selected from the group consisting of anesthetics, antihistamines, anticancer agents, antiulcers, antiseizure agents, muscle relaxants, immunosuppressants, anti-infective agents, nonsteroidal anti-inflammatory agents, imaging agents, nutrients, and combinations thereof. Can be. The active ingredient is methotrexate, doxorubicin, epirubicin, daunorubicin, vincristine, vinblastine, etoposide, elliptisin, camptothecin, paclitaxel, docetaxel, cisplatin, prednisone, methyl-prednisone, ibuprofen and combinations thereof It may be selected from the group consisting of.

Another aspect is a method of delivering an active ingredient to a target site in a subject, comprising administering to the subject a solid lipid nanoparticle comprising the active ingredient, wherein the solid lipid nanoparticle is conjugated to a moiety comprising a hydrophobic group. Comprising an elastin-like polypeptide and a lipid molecule; And heating the target site of the subject to release the active ingredient from the solid lipid nanoparticles at the target site.

The method comprises administering to the subject a solid lipid nanoparticle comprising the active ingredient. Solid lipid nanoparticles containing the active ingredient are as described above. The solid lipid nanoparticles may have a phase transition temperature of 39 ℃ to 45 ℃,

The administration can be parenteral administration. The parenteral administration can be administered, for example, by intravenous administration, subcutaneous, intramuscular, intraluminal (abdominal, joint, or intraocular), or direct injection. Direct injection may be direct injection at the site of the condition, eg, at the tumor site. The solid lipid nanoparticles can be administered in blood, such as veins, and delivered to a target site, such as a tumor site, by the bloodstream. The target site may have leaky properties. The phrase "leaky property" refers to a property with increased permeability of a substance as compared to normal tissue or cells. The target site may be a tumor site, in which the substance permeability of the blood vessels of the tumor is increased due to the leakyness of the tumor blood vessels.

The subject may be a human or a mammal other than a human. The mammal may be selected from the group consisting of dogs, cats, horses, cows, pigs, goats, monkeys, mice, and combinations thereof.

The method includes heating the target site of the subject to release the active ingredient from the solid lipid nanoparticles at the target site. The heating may be heated by a clinical process or may be associated with an inherently higher target site compared to other sites in the body such as inflammation. Heating by the clinical process may involve direct heat transfer, eg contacting the body with a bath of hot liquid medium, for example water, ultrasound at a target site, for example high intensity focused ultrasound. ) Or by applying a magnetic field, such as an amplified magnetic field (AMF), microwave and / or high frequency. The target site may be a location where a pathological condition exists, such as a tumor site (eg, a solid tumor), or a site where inflammation is present. The heating may be to heat to 39 ℃ to 45 ℃.

The active ingredient may be one or more selected from the group consisting of physiologically active agents, pharmaceutically active agents, magnetically active agents, imaging agents, and combinations thereof. The pharmaceutically active ingredient is selected from the group consisting of anesthetics, antihistamines, anticancer agents, antiulcers, antiseizure agents, muscle relaxants, immunosuppressants, anti-infective agents, nonsteroidal anti-inflammatory agents, imaging agents, nutrients, and combinations thereof. Can be. The active ingredient is methotrexate, doxorubicin, epirubicin, daunorubicin, vincristine, vinblastine, etoposide, elliptisin, camptothecin, paclitaxel, docetaxel, cisplatin, prednisone, methyl-prednisone, ibuprofen and combinations thereof It may be selected from the group consisting of.

According to the solid lipid nanoparticles according to one aspect, the dispersibility of the particles may be controlled by contraction or self-assembly of a elastin-like polypeptide conjugated to a moiety including a hydrophobic group according to temperature. Thus, the solid lipid nanoparticles can be used as a vehicle for efficiently delivering the active ingredient to the target site of the individual.

The solid lipid nanoparticles comprising the active ingredient, as well as the phase transition temperature of the solid lipid nanoparticles themselves, as well as the dispersibility and active component release properties of the particles by the phase transition temperature of the elastin-like polypeptide conjugated to the moiety containing a hydrophobic group This can be adjusted. Thus, when the solid lipid nanoparticles have a more stable composition at the body temperature, for example, molecules that stabilize lipid molecules such as cholesterol in an amount effective to maintain the solid lipid nanoparticles more stably at the body temperature. Even in the state in which the moiety containing the hydrophobic group is conjugated, the dispersibility of the particles and the release of the active ingredient can be efficiently controlled by the phase transition temperature of the elastin-like polypeptide.

According to another aspect, a pharmaceutical composition for delivering an active ingredient comprising a solid lipid nanoparticle to an individual may be used to efficiently deliver the active ingredient to the individual.

According to a method for administering an active ingredient to a target site in the body of an individual according to another aspect, the active ingredient can be efficiently delivered to a target site in the body of the individual.

FIG. 1 shows the water insoluble release profile of solid lipid nanoparticles comprising methyl red (MR), a poorly water soluble material prepared according to Example 1. FIG.
2 shows the size of solid lipid nanoparticles according to the amount of surfactant prepared according to Example 2. FIG.
FIG. 3 shows transmission electron micrographs of solid lipid nanoparticles (A) not containing coumarin-6 and solid lipid nanoparticles (B) including coumarin-6 prepared according to Example 3.
Figure 4 shows the size of the solid lipid nanoparticles according to the type of lipid molecules and the amount of surfactant prepared according to Example 4.
Figure 5 shows the presence of cholesterol oleate used as a stabilizer in the solid lipid nanoparticles prepared according to Example 7 and the size change of the particles with temperature.
FIG. 6 is a diagram showing the temperature sensitivity of drug ejection of solid lipid nanoparticles to which an elastin-like polypeptide encapsulated with paclitaxel according to Example 8 is introduced.
7 shows the results of intracellular delivery of coumarin-6 using solid lipid nanoparticles according to Example 9. FIG.

Hereinafter, the present invention will be described in more detail with reference to examples. However, these examples are for illustrative purposes only, and the scope of the present invention is not limited to these examples.

Example  One: Water solubility  Preparation of Solid Lipid Nanoparticles Containing Methyl Red

Solid lipid nanoparticles were prepared using lipid molecules. Mixing of chloroform and ethanol in a molar ratio of egg phosphatidylcholine (egg PC) as phospholipid, tricaprine and trilaurin as triglyceride (molar ratio of 6: 4), and cholesteryl oleate 5: 2: 1 as stabilizer Dissolve at room temperature and atmospheric pressure in a solvent (2: 1 volume ratio), add 1-2 ml of melt in a vessel containing 1 ml PBS and disperse by vortexing and sonication, evaporate the organic solvent by rotary evaporator, The solid lipid nanoparticles were formed in 1 ml of PBS because the organic solvent was evaporated (hereinafter referred to as "control SLN"). In addition, methyl red (MR), equivalent to 10% by weight of egg PC, was dissolved in a solvent with egg PC, cholesteryl oleate, and a mixture of tricaprine and trilaurin (molar ratio of 6: 4) as triglycerides. The MR-containing SLN was prepared in the same manner as described above except that it was referred to as "experimental group SLN".

1 shows the MR release profile of solid lipid nanoparticles prepared according to Example 1. FIG. FIG. 1 shows solid lipid nanoparticles containing poorly water-soluble substance (experimental group SLN) and solid lipid nanoparticles containing no water-soluble substance (control SLN) in 50 μL of PBS (SLN concentration of 1 mg of Egg PC (main lipid) ) / mL of PBS), and the absorbance value obtained over time at 410 nm, which is the maximum absorption wavelength of MR, while being stored at room temperature and atmospheric pressure. The horizontal axis represents absorbance measurement time, and the vertical axis represents absorbance value. As shown in FIG. 1, the release of MR over time was minimal at room temperature (less than 1% of the loading). That is, the solid lipid nanoparticles not having an elastin-like polypeptide had a very low degree of release of the encapsulated material.

In addition, the obtained solid lipid nanoparticles were measured for an average diameter using a Zetasizer Nano ZS (Malvern instrument, UK), a dynamic light scattering (DLS) instrument. Specifically, 10 μL of the solid lipid nanoparticles (1 mg of Egg PC / mL of PBS, 10 wt% MR of Egg PC) was added to a cuvette (Disposable solvent resistant MicroCuvette (ZEN0040), Malvern, UK) containing 90 μL of PBS, The diameter was measured by light scattering in Zetasizer. As a result, the particle size of the solid lipid nanoparticles containing methyl red was 168 nm ± 0.5 nm.

Example  2: Preparation of Solid Lipid Nanoparticles Including Surfactants

In this example, a mixed solvent of chloroform and ethanol in an egg phosphatidylcholine as phospholipid, a mixture of tricaprine and trilaurin as triglyceride (6: 4 molar ratio), and cholesteryl oleate in a molar ratio of 5: 2: 1 Dissolve at room temperature and atmospheric pressure in a: 1 volume ratio, and contain 1 ml PBS with 1 to 2 ml of the melt having different proportions (0, 2.5, 5, and 10 vol% of surfactant Tween 20) And dispersed by vortexing and sonication, and the organic solvent was evaporated by a rotary evaporator to form the solid lipid nanoparticles in 1 ml PBS because the organic solvent was evaporated.

The obtained solid lipid nanoparticles were measured for average diameters using a Zetasizer Nano ZS (Malvern instrument, UK), a dynamic light scattering (DLS) instrument. Specifically, 10 μL of the solid lipid nanoparticles (1 mg of Egg PC / mL of PBS, 10 wt% MR of Egg PC) was added to a cuvette (Disposable solvent resistant MicroCuvette (ZEN0040), Malvern, UK) containing 90 μL of PBS, The diameter was measured by light scattering in Zetasizer. As a result, the resulting solid lipid nanoparticles had an average diameter of about 50 nm to 650 nm.

2 shows the size of solid lipid nanoparticles according to the amount of surfactant prepared according to Example 2. FIG. As shown in FIG. 2, the particle size decreased as the amount of Tween 20 increased.

Example  3: Water solubility  Preparation of Solid Lipid Nanoparticles Containing Materials and Identification of Transmission Electron Microscopy

In this example, egg phosphatidylcholine as a phospholipid, a mixture of tricaprine and trilaurin as a triglyceride (6: 4 molar ratio), and cholesteryl oleate in a molar ratio of 5: 2: 1, coumarin- which is a poorly water-soluble substance. 6 (10% by weight of phosphatidylcholine) and simultaneously dissolved in a mixed solvent of chloroform and ethanol (2: 1 volume ratio) at room temperature and atmospheric pressure, and 1-2 ml of the melt was dissolved in 1 ml PBS (2 volume% Tween 20 based on PBS volume). In) and dispersed by vortexing and sonication, and the organic solvent was evaporated by a rotary evaporator to form the solid lipid nanoparticles in 1 ml PBS.

The obtained solid lipid nanoparticles were measured for average diameters using a Zetasizer Nano ZS (Malvern instrument, UK), a dynamic light scattering (DLS) instrument. Specifically, 10 μL of the solid lipid nanoparticles (1 mg of Egg PC / mL of PBS, 10 wt% coumarin-6 of Egg PC) was added to a cuvette (Disposable solvent resistant MicroCuvette (ZEN0040), Malvern, UK) containing 90 μL of PBS. And the diameter was measured by light scattering in Zetasizer. As a result, the resulting solid lipid nanoparticles had an average diameter of about 200 nm to 220 nm. In addition, when observed using a transmission electron microscope, it was confirmed that coumarin-6 particles were introduced into the solid lipid molecule.

3 shows transmission electron micrographs of solid lipid nanoparticles (A) not containing coumarin-6 and solid lipid nanoparticles (B) including coumarin-6. As shown in FIG. 3B, black solids corresponding to coumarin-6 were present in the solid lipid nanoparticles.

In addition, the stability of the produced solid lipid nanoparticles was investigated. Solid lipid nanoparticles prepared according to Example 1, a solution of 1 mg of Egg PC / ml of PBS (without MR) concentration for 1 week at room temperature and atmospheric pressure, the same as above after 24 hours and 72 hours on the day of preparation The particle size was measured by means of Zetasizer. The average particle size was 168 nm ± 0.5 nm and there was no change in particle size even when stored for about 72 hours.

Example  4: Preparation of solid lipid nanoparticles according to the type of lipid molecules and the amount of surfactant

In the present Example, solid lipid nanoparticles were prepared in the same manner as in Example 3, but the phospholipid type was phosphatidylcholine or dipalmitoylphosphatidylcholine (DPPC), and triglyceride ratio tricarphrine to trilaurin was used. Was changed to 8: 2 to prepare lipid nanoparticles (coumarin-6 free). In addition, the amount of Tween 20 was used as 0,1, or 2% by volume of PBS. The prepared lipid nanoparticle concentration was 1 mL of 1 mg / Tween 20 containing PBS.

The obtained solid lipid nanoparticles were measured for average diameters using a Zetasizer Nano ZS (Malvern instrument, UK), a dynamic light scattering (DLS) instrument. Specifically, 10 μL of the solid lipid nanoparticles (1 mg of Egg PC / mL of PBS) was added to a cuvette (Disposable solvent resistant MicroCuvette (ZEN0040), Malvern, UK) containing 90 μL of PBS, and the diameter was determined by light scattering in Zetasizer. Measured.

Figure 4 shows the size of the solid lipid nanoparticles according to the type of lipid molecules and the amount of surfactant. 4 is a solution prepared by mixing 10 μL of a solid lipo-nanoparticle solution prepared in the same manner as in Example 4 and 90 μL of PBS, and is maintained at 25 ° C. and 45 ° C. for 5 minutes, using Zetasizer as described above. This is the result of measuring the average diameter. As shown in FIG. 4A, the resulting solid lipid nanoparticles had an average diameter of about 200 nm to about 950 nm at 25 ° C. and about 50 nm to about 400 nm at 45 ° C. when the phospholipid PC was PC. In addition, as shown in FIG. 4B, when the phospholipid was DPPC, the average diameter was about 500 nm to about 2580 nm at 25 ° C., and about 250 nm to about 700 nm at 45 ° C. FIG.

Example  5: Preparation and Particle Size Measurement of Solid Lipid Nanoparticles Comprising Elastin-Like Polypeptides

Stearoyl- (VPGVG) n-NH _{2 with} n of 1 to 200 as an elastin-like polypeptide (Hereinafter SA-Vn) This was used to prepare solid lipid nanoparticles. Here, the stearoyl group is linked by an N-terminal nitrogen and an amide bond.

In this example, SA-V5 and SA-V6 (peptron, Korea) (n weight 5 and 6) (n weight 10% by weight of PC), egg phosphatidylcholine as phospholipid, a mixture of tricaprine and trilaurin as triglycerides (6: 4 mole ratio) and cholesteryl oleate in a molar ratio of 5: 2: 1 (I PC: triglyceride: cholesteryl oleate) in a mixed solvent of chloroform and ethanol (2: 1 volume ratio) at room temperature and atmospheric pressure, The organic solvent was evaporated by rotary evaporator to film the lipid solution. PBS is added to obtain a solid lipid nanoparticle solution in which particles are dispersed by vortexing and sonication.

The obtained solid lipid nanoparticles were measured for average diameters using a Zetasizer Nano ZS (Malvern instrument, UK), a dynamic light scattering (DLS) instrument. Specifically, 10 μL of the solid lipid nanoparticles (1 mg of Egg PC / mL of PBS, 10 wt% ELP of Egg PC) was added to a cuvette (Disposable solvent resistant MicroCuvette (ZEN0040), Malvern, UK) containing 90 μL of PBS, The diameter was measured by light scattering in Zetasizer. As a result, the size of the solid lipid nanoparticles produced was 220 nm (SA-V5) and 120 nm (SA-V6), depending on the length of the elastin-like polypeptide used.

Example  6: Preparation of Solid Lipid Nanoparticles Comprising Elastin-Like Polypeptides and Particle Size Variation with Temperature

Cholesteryl- (VPGVG) n1- (VPGAG) n2-NH ₂ (hereinafter referred to as Chol-Vn1An2) as elastin-like polypeptides, wherein n1 and n2 are 1 to 200, respectively. Was prepared.

In this example, Chol-Vn1An2 (n, 0, 1, 2.5, and 5 wt% of phosphatidylcholine) (peptrone, Korea), n1 is 3 and n2 is 1, phosphatidylcholine as phospholipid, tricaprine and trilaurin as triglyceride Mixture of (8: 2 molar ratio), and cholesteryl oleate at room temperature in a mixed solvent of chloroform and ethanol (2: 1 volume ratio) at a molar ratio of 5: 2: 1 (I PC: triglyceride: cholesteryl oleate). And dissolved at atmospheric pressure, and the organic solvent was evaporated by rotary evaporator to film the solid lipid nanoparticles. PBS is added to obtain a solid lipid nanoparticle solution in which particles are dispersed by vortexing and sonication.

The obtained solid lipid nanoparticles were measured for average diameter using Zetasizer Nano ZS (Malvern instrument, UK), a dynamic light scattering (DLS) instrument. Specifically, 10 μL of the solid lipid nanoparticles (1 mg of Egg PC / mL of PBS, 0, 1, 2.5, and 5 wt% ELP of Egg PC) and 90 μL of PBS (Disposable solvent resistant MicroCuvette (ZEN0040), Malvern, UK) and measured the diameter by light scattering in Zetasizer.

Tables 1 and 2 show the sizes of the solid lipid nanoparticles measured as described above.

Name of sample Main peak (d.nm) 2nd peak (d.nm) 3rd peak (d.nm) PdI w / o Chol-V3A-25 ℃ 1046 (100%) - - 0.498 1% Chol-V3A-25 ℃ 326.9 (61%) 82.92 (37%) 5204 (2%) 0.412 2.5% Chol-V3A-25 ℃ 181 (90%) 34.22 (6%) 5194 (4%) 0.379 5% Chol-V3A-25 ℃ 212.3 (98%) 5207 (2%) - 0.4

Name of sample Main peak (d.nm) 2nd peak (d.nm) 3rd peak (d.nm) PdI w / o Chol-V3A-42 ℃ 134.2 (67%) 1746 (33%) - 0.677 1% Chol-V3A-42 ℃ 178.4 (100%) - - 0.296 2.5% Chol-V3A-42 ℃ 148.1 (100%) - - 0.138 5% Chol-V3A-42 ℃ 164.7 (100%) - - 0.304

*%: Percent of particle of the corresponding size in the total particle distribution

** Pdi: polydispersity index

*** d: diameter

As shown in Tables 1 and 2, the resulting solid lipid nanoparticles, when comprising elastin-like polypeptides, had an average diameter of about 180 nm to 330 nm at 25 ° C. and about 150 nm to 180 nm at 42 ° C. In the absence of an elastin-like polypeptide, the average diameter was 25 ° C. at about 1050 nm and 45 ° C. at about 134 nm to 1750 nm. The introduction of the elastin-like polypeptide Chol-V3A into the solid lipid nanoparticles was confirmed to increase the temperature sensitivity of the particles. When containing an elastin-like peptide, not only the particle size decreases with temperature, but also the dispersion of the particles was improved. For example, the degree of dispersion decreased the Pdi value at 45 ° C than at 25 ° C.

Example  7: Preparation and central portion of solid lipid nanoparticles comprising an elastin-like polypeptide core lipid Changes in Particle Size with Components and Temperature

Cholesteryl- (VPGVG) n1 (VPGAG) n2-NH ₂ (hereinafter referred to as Chol-Vn1An2): VPGVG (SEQ ID NO: 6) and VPGAG (SEQ ID NO: 7) as elastin-like polypeptides Solid lipid nanoparticles were prepared.

In this example, Chol-Vn1An2 (1 wt% of egg PC), n1 is 3 and n2 is 1, egg phosphatidylcholine as phospholipid, a mixture of tricaprine and trilaurin as triglyceride (7: 3 molar ratio), and cholesteryl The oleate is dissolved in a mixed solvent of chloroform and ethanol (2: 1 volume ratio) at room temperature and atmospheric pressure in a molar ratio of 5: 2: 1 or 5: 2: 0, and the organic solvent is evaporated by a rotary evaporator to give the solid lipid nano The particles were filmed. To the obtained solid lipid nanoparticle film, PBS (with or without 5 vol% triton X-100 based on PBS volume: 50 μL when 1 mL of PBS was added) was added, and particles were dispersed by vortexing and sonication. A solid lipid nanoparticle solution is obtained. Triton X-100 is a surfactant and has the same nonionic properties as Tween.

The obtained solid lipid nanoparticles were measured for average diameter using Zetasizer Nano ZS (Malvern instrument, UK), a dynamic light scattering (DLS) instrument. Specifically, 10 μL of the solid lipid nanoparticles (1 mg of Egg PC / mL of PBS, 1 wt% ELP of Egg PC) was added to a cuvette (Disposable solvent resistant MicroCuvette (ZEN0040), Malvern, UK) containing 90 μL of PBS. The diameter was measured by light scattering in Zetasizer.

FIG. 5 shows the temperature sensitivity of particle dispersion depending on whether or not cholesteryl oleate, which acts as a stabilizer of solid lipid nanoparticles, is introduced into the core of the particle, according to Example 7. As shown in FIG. 5, the solid lipid nanoparticles, when including cholesteryl oleate, had an average diameter of about 718 nm at 25 ° C., about 236 nm at 37 ° C., and about 206 nm at 42 ° C. FIG. In addition, the solid lipid nanoparticles, when not containing cholesteryl oleate, had an average diameter of about 630 nm at 25 ° C, 210 nm at 37 ° C, and about 190 nm at 42 ° C. When cholesteryl oleate, which acts as a stabilizer of SLN, was introduced into the core of the particles, it was confirmed that the temperature sensitivity and dispersibility of the solid lipid nanoparticles having elastin-like polypeptides were increased.

Example  8: of solid lipid nanoparticles comprising an elastin-like polypeptide Parkle Retaxel filling and temperature-dependent release confirmation

Cholesteryl- (VPGVG) n1 (VPGAG) n2-NH ₂ with n of 1 to 200 as ELP (Hereinafter referred to as Chol-Vn1An2) to prepare solid lipid nanoparticles.

In this example, Chol-Vn1An2 (1 wt% of egg PC) (Peptron, Korea), n-phosphate 3, n2 is 1, paclitaxel (10% by weight of egg PC) and egg phosphatidylcholine as phospholipid and tricaprine as triglyceride. A mixture of trilaurin (7: 3 molar ratio) (I PC: TG = 5: 2 molar ratio) is dissolved in a mixed solvent of chloroform and ethanol (2: 1 volume ratio) at room temperature and atmospheric pressure, and the organic solvent is dissolved by a rotary evaporator. The solid lipid nanoparticles were filmed by evaporation. PBS is added to obtain a solid lipid nanoparticle solution in which particles are dispersed by vortexing and sonication.

The obtained solid lipid nanoparticles were measured for average diameters using a Zetasizer Nano ZS (Malvern instrument, UK), a dynamic light scattering (DLS) instrument. Specifically, 10 μL of the solid lipid nanoparticles (1 mg of Egg PC / mL of PBS, 10 wt% paclitaxel of Egg PC) was added to a cuvette (Disposable solvent resistant MicroCuvette (ZEN0040), Malvern, UK) containing 90 μL of PBS, The diameter was measured by light scattering in Zetasizer. When measured in Zetasizer, the average diameter was measured while maintaining at 25 ° C and 42 ° C for 5 minutes.

As a result, when the solid lipid nanoparticles containing paclitaxel included Chol-V3A, the average diameter was about 720 nm at 25 ° C, and about 240 nm at 42 ° C. Without Chol-V3A, the average diameter was about 1350 nm at 25 ° C. and about 310 nm at 42 ° C. As a result, when Chol-V3A is introduced into the solid lipid nanoparticles, it was confirmed that the dispersibility of the particles increases with increasing temperature. When measured on the Zetasizer, the PDI value obtained was reduced and the average size of the particles measured in a given sample was reduced, which is believed to indicate that the size difference between the particles was reduced.

FIG. 6 is a diagram showing the temperature sensitivity of drug release of solid lipid nanoparticles to which an elastin-like polypeptide encapsulated with paclitaxel according to Example 8 is introduced. As shown in FIG. 6, the drug was rapidly released at about 20 minutes at 42 ° C., but similar release tendency was observed at 25 ° C. and 37 ° C., and there was no rapid release even after about 60 minutes.

6 is a result of confirming the degree of ejection of paclitaxel according to the temperature from the solid lipid nanoparticles using a semipermeable membrane and UV. Specifically, 2.5 mg (5 mg of egg PC / ml) of solid lipid nanoparticles were added to a semi-permeable membrane (Spectra / Por ^TM Dialysis membrane, MWCO 1,000) in 25 mL PBS (1 vol% Tween 20) medium, 25 ° C., 37 incubated at ℃ and 42 ℃, and semi-permeable membrane at a specified time and the absorbance of the solution was measured at the outside paclitaxel maximum absorption wavelength (λ _max) 240nm. As a result, more than twice as much drug was released at 42 degreeC over 10 minutes compared with 25 degreeC and 37 degreeC.

Example  9: coumarin-6 inclusion and temperature dependence of solid lipid nanoparticles comprising elastin-like polypeptides Intracellular  relay ( cellular uptake ) Confirm

Cholesteryl- (VPGVG) n1 (VPGAG) n2-NH ₂ (hereinafter referred to as Chol-Vn1An2) with elastin-like polypeptides was used to prepare solid lipid nanoparticles.

In this example, Chol-Vn1An2 (1 wt% of egg PC), coumarin-6 (10 wt% of egg PC amount), n1 is 3 and n2 is 1, and phosphatidylcholine as triphosphide and tricaprine and trila as triglycerides Our mixture (7: 3 molar ratio) (I PC: TG = 5: 2 molar ratio) is dissolved in a mixed solvent of chloroform and ethanol (2: 1 volume ratio) at room temperature and atmospheric pressure, and the organic solvent is evaporated by rotary evaporator. To solidify the lipid lipid nanoparticles. PBS is added to obtain a solid lipid nanoparticle solution in which particles are dispersed by vortexing and sonication.

In order to confirm the degree of intracellular delivery of the obtained solid lipid nanoparticles, it was analyzed by flow cytometry. Solid lipid nanoparticles containing coumarin-6 were treated with KB cells, which are cancer cells, and after 2 hours, the degree of intracellular delivery was measured using fluorescence (using a FITC filter). Specifically, KB cells (ATCC Number: CCL-17 ^™ ) were grown in a 12-well plate containing 500ul of growth medium (10% (v / v) FBS containing ATCC-formulated Eagle's Minimum Essential Medium) of 2.5 × 10 ⁵ cells / well. Each well was inoculated at a concentration and incubated in a 37 ° C., 5% CO ₂ incubator until the cell concentration was 80% confluence. To each well's medium, add 500 μl of the fresh growth medium and add 9 μg / ml of coumarin-6 per well to it, and leave it at 37 ° C. or 42 ° C. for 10 minutes, then 37 ° C., 5% CO ₂ for 2 hours. After incubation in the incubator, flow cytometry was performed using a fluorescence-activated cell sorting (FACS) (Canto II, BD bioscience) instrument to measure fluorescence emitted from the cells.

7 shows the results of intracellular delivery of coumarin-6 using solid lipid nanoparticles according to Example 9. FIG. As shown in FIG. 7, when coumarin-6, a poorly water-soluble substance, was enclosed in solid lipid nanoparticles, intracellular delivery increased by 100 times or more. In FIG. 7, SLN-1 represents solid lipid nanoparticles without Chol-V3A and SLN-4 represents solid lipid nanoparticles with Chol-V3A. Control indicates coumarin-6 treatment directly to cells (treatment in 12 well plates at a concentration of 9 μg / ml, 4.5 μg of coumarin-6 per well). In FIG. 7, the number of cells showed that the solid lipid nanoparticles were internalized in more cells in SNL4 than in SLN1. In FIG. 7, the cell number is assessed as the area value under the graph, and for SLN-1, the range of X values was about 5,000 to 30,000, which was smaller than the range of SLN-4, which is about 9,000 to 60,000.

Example  10: multiple Acyl  Solid lipid nanoparticles comprising an elastin-like polypeptide having

Solid lipid nanoparticles were prepared using SA- [K '(SA) V' ₃ ] ₂ -NH ₂ as an elastin-like polypeptide. Here, SA is stearoyl, K 'represents VPGKG (SEQ ID NO: 8), K' (SA) represents that the stearoyl group is conjugated to NH ₂ of the lysine side chain of VPGKG, V 'represents VPGXG, One of PGXGV, GXGVP, XGVPG, and GVPGX, and the 1st, 2nd, 3rd V ′ may be the same or different sequences. Here, V may be valine, P is proline, G is glycine, and X may be a natural or non-natural amino acid other than proline. Here, X of each repeating unit may be the same or different amino acids.

In this embodiment, SA-[(VPGK (SA) G) (VPGVG) ₃ ] ₂ -NH ₂ (SEQ ID NO: 9, 3wt% of egg PC), paclitaxel (10% by weight of egg PC), egg phosphatidylcholine as phospholipid in a 5: 2: 1 molar ratio, a mixture of tricaprine and trilaurin as triglycerides (7: 3 Molar ratio), and cholesterol oleate in a mixed solvent of chloroform and ethanol (2: 1 volume ratio) at room temperature and atmospheric pressure, and the organic solvent was evaporated by rotary evaporator to film the solid lipid nanoparticles. PBS containing 1% by volume Tween 20 was added and the vortexing and sonication were repeated to obtain a solution of solid lipid nanoparticles sprayed with particles (SLN 5 mg / ml). As a control, SLN without cholesterol oleate was prepared by the same procedure.

The obtained solid lipid nanoparticles were measured for average diameters using a Zetasizer Nano ZS (Malvern instrument, UK), a dynamic light scattering (DLS) instrument. Specifically, 10 μL of the solid lipid nanoparticles (1 mg of Egg PC / mL of PBS, 10 wt% paclitaxel of Egg PC) was added to a cuvette (Disposable solvent resistant MicroCuvette (ZEN0040), Malvern, UK) containing 90 μL of PBS, The diameter was measured by light scattering in Zetasizer.

As a result, the SLN particle size was about 360 nm and 310 nm at 37 ° C. and 42 ° C., respectively, when containing cholesterol oleate (hereinafter referred to as “experimental group”), and when not including oleate (hereinafter referred to as “control”. ), 37 ° C. and 42 ° C. at about 270 nm and 270 nm, respectively. That is, in the case of containing oleate, the size of SLN was small when the temperature rose to 37 ° C or higher. When measured in Zetaziser, the average diameter was measured while maintaining at 37 ° C and 42 ° C for 5 minutes.

Table 3 shows the measured dispersion of SLN with temperature.

Name of sample Polydispersity index 25 ℃ 37 ℃ 42 ° C Experimental group 0.63 0.14 0.24 Control group 0.44 0.24 0.23

As shown in Table 3, the polydispersity coefficient of SLN decreased with increasing temperature in both experimental and control groups. That is, as the temperature increased, the SLN in 1 wt% tween 20 containing PBS became a more uniformly dispersed solution. Accordingly, it is expected that paclitaxel included in SLN will be easier to release. As the degree of dispersion increases, the larger particles are divided into smaller particles, thereby increasing the surface area of the solid lipid nanoparticles and exposing the drug introduced therein to the outside of the lipid particles, i.e., the solvent. It is therefore expected that the release of the drug will be facilitated as the particles become smaller and the degree of dispersion increases.

<110> Samsung Electronics Co., Ltd. <120> Solid lipid nanoparticles including elastin-like polypeptide and          use thereof <130> PN0100102 <160> 9 <170> Kopatentin 2.0 <210> 1 <211> 5 <212> PRT <213> Artificial Sequence <220> <223> elastin-like polypeptide unit sequence <220> <221> VARIANT <222> (4) <223> Xaa denotes amino acid other than proline <400> 1 Val Pro Gly Xaa Gly   1 5 <210> 2 <211> 5 <212> PRT <213> Artificial Sequence <220> <223> elastin-like polypeptide unit sequence <220> <221> VARIANT <222> (3) <223> Xaa denotes amino acid other than proline <400> 2 Pro Gly Xaa Gly Val   1 5 <210> 3 <211> 5 <212> PRT <213> Artificial Sequence <220> <223> elastin-like polypeptide unit sequence <220> <221> VARIANT <222> (2) <223> Xaa denotes amino acid other than proline <400> 3 Gly Xaa Gly Val Pro   1 5 <210> 4 <211> 5 <212> PRT <213> Artificial Sequence <220> <223> elastin-like polypeptide unit sequence <220> <221> VARIANT <222> (1) <223> Xaa denotes amino acid other than proline <400> 4 Xaa Gly Val Pro Gly   1 5 <210> 5 <211> 5 <212> PRT <213> Artificial Sequence <220> <223> elastin-like polypeptide unit sequence <220> <221> VARIANT <222> (5) <223> Xaa denotes amino acid other than proline <400> 5 Gly Val Pro Gly Xaa   1 5 <210> 6 <211> 5 <212> PRT <213> Artificial Sequence <220> <223> Elastin-like polypeptide <400> 6 Val Pro Gly Val Gly   1 5 <210> 7 <211> 5 <212> PRT <213> Artificial Sequence <220> <223> Elastin-like polypeptide <400> 7 Val Pro Gly Ala Gly   1 5 <210> 8 <211> 5 <212> PRT <213> Artificial Sequence <220> <223> Elastin-like polypeptide unit sequence <400> 8 Val Pro Gly Lys Gly   1 5 <210> 9 <211> 40 <212> PRT <213> Artificial Sequence <220> Elastin-like polypeptide unit sequence; the nitrogens at the          N-terminal end and lysine side chains are conjugated to stearoyl          groups and C-terminal end is modified to -NH2 <400> 9 Val Pro Gly Lys Gly Val Pro Gly Val Gly Val Pro Gly Val Gly Val   1 5 10 15 Pro Gly Val Gly Val Pro Gly Lys Gly Val Pro Gly Val Gly Val Pro              20 25 30 Gly Val Gly Val Pro Gly Val Gly          35 40

Claims (13)

Solid lipid nanoparticles comprising an elastin-like polypeptide (ELP) conjugated with a moiety comprising at least one hydrophobic group and a lipid molecule, wherein the moiety comprising the hydrophobic group is a saturated or unsaturated hydrocarbon group, A saturated or unsaturated acyl group, R 1 and R 2 are each independently a saturated or unsaturated hydrocarbon, a substituted amide group or a saturated or unsaturated alkoxy group of formula -C (CO) N (R 1) (R 2), wherein the lipid molecule is a neutral lipid Molecule, amphipathic lipid molecule, or a combination thereof, wherein the elastin-like polypeptide comprises one or more repeating units selected from the group consisting of VPGXG, PGXGV, GXGVP, XGVPG, GVPGX, and combinations thereof, wherein V Is valine, P is proline, G is glycine, and X is a non-proline amino acid. The solid lipid nanoparticle of claim 1, wherein the moiety including the hydrophobic group has two or more of the moieties conjugated to one elastin-like polypeptide molecule. The solid lipid nanoparticle of claim 1, wherein the moiety including the hydrophobic group has at least one moiety conjugated to a side chain of an elastin-like polypeptide molecule. The solid lipid nanoparticle of claim 1, further comprising a stabilizer. The solid lipid nanoparticles of claim 1, wherein the lipid molecules constituting the solid lipid nanoparticles include those having a phase transition temperature of 39 ° C to 60 ° C. The solid lipid nanoparticle of claim 1, having an average diameter of about 10 nm to about 1500 nm. The solid lipid nanoparticle of claim 1, further comprising at least one selected from the group consisting of physiologically active agents, pharmaceutically active agents, magnetically active agents, imaging agents, and combinations thereof. The elastin-like polypeptide conjugated to a moiety comprising at least one hydrophobic group, a first lipid molecule, a second lipid molecule, and a stabilizer; wherein the first lipid molecule is a phospholipid and the second lipid molecule. Is a solid lipid nanoparticle which is a neutral lipid molecule, wherein the stabilizing agent is selected from the group consisting of sterols or derivatives thereof, sphingolipids or derivatives thereof, and combinations thereof, wherein the phospholipids have C16-C24 acyl groups, The neutral lipid molecule is a solid lipid nanoparticle comprising at least one of monoglycerides, diglycerides and triglycerides of at least one C4 to C24 carboxylic acid. The moiety of claim 8 comprising a elastin-like polypeptide conjugated to a moiety comprising at least one hydrophobic group, a triglyceride consisting of tricaprine and trilaurin and cholesteryl oleate, wherein the moiety comprising said at least one hydrophobic group. The tee conjugated elastin-like polypeptide has n between 1 and 200 and V ′ represents any one of VPGXG, PGXGV, GXGVP, XGVPG, and GVPGX, and when n is 2 or more, V ′ at each position in the sequence is the same or equal to Other sequences, V is valine, P is proline, G is glycine, X is a natural or non-natural amino acid that is not proline, and X 'at each position is the same or a different amino acid stearoyl- or cholesteryl -V'n-NH ₂ ;
n ₁ , n ₂ and n ₃ are each 1 to 200 and V ₁ And V ₂ represents any one of VPGXG, PGXGV, GXGVP, XGVPG, and GVPGX, and when n ₁ and n ₂ are each 2 or more, V ′ at each position in the sequence is the same or different sequence, and V is valine, P Is proline, G is glycine, X is a non-proline natural or non-natural amino acid, and X at each position of V 'is the same or a different amino acid, such as stearoyl- or cholesteryl- [V ₁ n ₁ V ₂ n ₂ ] n ₃ -NH ₂ ; or
n ₁ , n ₂ and n ₃ are each from 1 to 200, and B (SA or Chol) is any one of VPGXG, PGXGV, GXGVP, XGVPG, and GVPGX, where V is valine, P is proline, G is glycine, X represents a lysine, arginine or histidine with a stearoyl- or cholesteryl-conjugated side chain amino group, V ₁ is any of VPGXG, PGXGV, GXGVP, XGVPG, and GVPGX, where V is valine and P is proline , G represents glycine, X represents a natural or non-natural amino acid that is not proline, and when n ₁ or n ₂ is a number of two or more, B and V ₁ at each position are the same or different sequences, respectively, stearoyl- or chole Solid lipid nanoparticles which are steryl- [B (SA or Chol) n ₁ V ₁ n ₂ ] n ₃ -NH ₂ . 10. The molar ratio of phosphatidylcholine, triglyceride and cholesteryl oleate consisting of tricaprine and trilaurine is about 2-5: molar ratio of about 0.1-3: about 0-1, and tricaprine and tri The molar ratio of laurin is from 1: 0.25 to about 4. The elastin-like polypeptide conjugated to the moiety comprising at least one hydrophobic group is from about 0.01 to about 50 weight percent of egg PC. A pharmaceutical composition for delivering to an individual an active ingredient comprising a solid lipid nanoparticle according to any one of claims 1 to 10 comprising a pharmaceutically acceptable carrier or diluent, and the active ingredient, wherein the active ingredient is a physiological A composition comprising one or more selected from the group consisting of an active agent, a pharmaceutically active agent, a magnetically active agent, an imaging agent, and a combination thereof. A method of delivering an active ingredient to a target site in an individual's body,
Administering to the individual a solid lipid nanoparticle according to any one of claims 1 to 10 comprising the active ingredient; And
Heating the target site of the subject to release the active ingredient from the solid lipid nanoparticles at the target site, wherein the active ingredient is a physiologically active agent, a pharmaceutically active agent, a magnetically active agent, imaging And at least one selected from the group consisting of a combination thereof. The method of claim 12, wherein the heating is heated to 39 ° C. to 45 ° C.

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