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Multilamellar lipid vesicle compositions and methods of use

A technology of lipid vesicles and compositions, which is applied in the field of multilamellar lipid vesicle compositions and uses, can solve problems such as high risks, and achieve the effect of improving encapsulation efficiency

Inactive Publication Date: 2018-03-27
VEDANTRA PHARMA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, by the time HPV infection has manifested in one or more of these symptoms, the risk of developing HPV-related cancers, such as cancers of the cervix, anus, vagina, vulva, penis, or oropharynx, becomes extremely high

Method used

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  • Multilamellar lipid vesicle compositions and methods of use
  • Multilamellar lipid vesicle compositions and methods of use
  • Multilamellar lipid vesicle compositions and methods of use

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0346] Example 1. Synthesis of ICMV and Conjugated Reagents

[0347] Material:

[0348] (a) DOPC (5mg / ml in CHCl 3 Medium - Avanti Polar Lipids #850375)

[0349] (b) MPB (10mg / ml in CHCl 3 Medium - Avanti Polar Lipids #870012)

[0350] (c) 3M-052 (0.1mg / ml in CHCl 3 Medium) or MPLA / GLA (0.5mg / ml in MeOH)

[0351] (d) (PEG 2k)-SH (100mg / ml in H 2 Om-Laysan Bio#MPEG-SH-2000)

[0352] (e) dithiothreitol (150mM in H 2 O Medium - Sigma Aldrich #43819)

[0353] (f) CaCl 2 (200mM in H 2 O Medium - Fischer Scientific #BP510)

[0354] (g) Reagents at appropriate concentrations in 20 mM bis-bis(tris-methylol)methylaminopropane (bis-Tris-propane), pH 7.0 (bTp-Sigma Aldrich #B4679)

[0355] (h) Trout's reagent (10 mg / ml in bTp - Thermo Scientific #26101)

[0356] program:

[0357] DOPC (500 μL), MPB (325 μL), and optional adjuvant components were dried under vacuum in scintillation vials for at least 12 hours, followed by vortexing at room temperature at 3000 rpm for 10 seco...

Embodiment 2

[0362] Example 2. Optimization of Reagent Functionalization

[0363] Ovalbumin with different molar equivalents of Trout's reagent resulted in different functionalization ratios. The amount of free thiols was determined using Elman's analysis.

[0364] Material:

[0365] Ellman's reagent (Pierce); Ellman's reaction buffer is 0.1 M sodium phosphate, 1 mM EDTA, pH 8.0; cysteine ​​hydrochloride monohydrate (Pierce)

[0366] Program:

[0367] Ellman's reagent (4 mg) was dissolved in Ellman's reaction buffer (1 mL). Cysteine ​​standards and samples were added to 96-well plates in triplicate at 20 uL / well. Ellman's reagent (20 μL per well) and PBS (60 μL per well) were added and incubated for 15 minutes. Absorbance was read at a fixed wavelength of 412 nm (reference wavelength 540 nm). The results are shown in Table 2.

[0368] Table 2. Functionalization of Ovalbumin

[0369]

[0370] Ovalbumin has an average of 0.35 molecules of added sulfur per molecule when it is ...

Embodiment 3

[0375] Example 3. Optimization of Reagent Conjugation

[0376] Experiments were performed to determine whether increasing the time for reagents to be conjugated to the fluidized lipids would increase encapsulation efficiency. The results are shown in Table 4.

[0377] Table 4. Optimization of Reagent Conjugation Time

[0378] sample

[0379] Due to the high reactivity of the maleimide groups on ICMVs with free thiol groups, the reaction of thiol-containing reagents with ICMVs is usually limited to 1 h, but since functionalization of large protein molecules is slow, it was found to provide More time for this reaction, an increase in protein loading efficiency of up to 47% occurred. Thus, the desired encapsulation efficiency is achieved during the 24 hour incubation period, resulting in increased conjugation of the reagents.

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Abstract

The present invention provides novel and inventive drug delivery systems with higher loading capability, a capacity to sequester high levels of both hydrophobic and hydrophilic agents simultaneously,and longer release profiles. Some aspects of these delivery systems include compositions including stabilized multilamellar lipid vesicles having crosslinked lipid bilayers (referred to herein as interbilayer-crosslinked multilamellar vesicles or ICMV) covalently conjugated to an agent (e.g., an antigen).

Description

Background of the invention [0001] Liposomes have been widely used as delivery vehicles for small molecules; however, it is still difficult to achieve a high level of encapsulation of many macromolecular drugs within liposomes, and many drug formulations leak from liposomes too quickly to keep Useful drug delivery kinetics. While drug delivery via micro- and nanoparticles can encapsulate proteins and small molecule drugs, this still typically results in very low total mass encapsulated drug per unit mass of particle, typically on the order of about 10 μg drug / mg particle. Furthermore, the organic solvents used for polymer particle synthesis and the hydrophobic / acidic environment within these particles can lead to the destruction of therapeutic agents (see Zhu et al. Nat. Biotechnol. 2000 18:52-57). [0002] One area that could benefit from efficient drug delivery via micro- and nanoparticles is human papillomavirus (HPV)-associated cancer. HPV-related cancers are among the f...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K31/56A61K9/127
CPCC07K2319/00C12N2710/20034A61K39/12A61K31/56A61K38/162A61P31/00A61P35/00A61P35/02A61K39/001193A61K39/001194A61K39/00117A61K39/001191A61K39/00115A61K39/001166A61K39/001164A61K39/001182A61K39/001197A61K39/001156A61K39/001192A61K39/001104A61K39/001152A61K39/001184A61K39/001122A61K39/001188A61K39/001106A61K39/001151A61K39/001172A61K39/001181A61K39/001109A61K39/001149A61K39/00119A61K39/0011A61K39/001108A61K39/001153A61K39/001168A61K39/001171A61K39/001186A61K39/001195A61K35/76A61K2039/525A61K2300/00C07K16/30C12N2710/00032
Inventor 李赛君J.埃比P.C.德穆思
Owner VEDANTRA PHARMA
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