Extract and formulation including extract

A method for making a preparation containing an extract from inflamed skins of rabbits inoculated with vaccinia virus, comprising providing an extract from inflamed skins of rabbits inoculated with vaccinia virus having a predetermined amount of N-acetylneuraminic acid where the variations in the content of N-acetylneuraminic acid among manufacturing lots are reduced, the amount of N-acetylneuraminic acid contained in the extract being 4000 ng or more per unit of the extract, and incorporating the extract into an injectable preparation or a tablet preparation,

• wherein the extract is obtained by a manufacturing method comprising:
• (A) collecting inflamed skin tissues of rabbits subjected to the intradermal inoculation with vaccinia virus, and crushing the inflamed tissues, conducting an extraction treatment comprising adding to the crushed tissues an extraction solvent in 1- to 5-fold thereof by volume of water, phenol water, physiological saline or phenol-added glycerin water to give a mixture, allowing the mixture to stand for 3 to 7 days and further heating the mixture at 35 to 40° C., then, filtrating or centrifuging the mixture to give a crude extract which is a filtrate or supernatant, and removing tissue fragments from the crude extract,
• (B) adjusting the crude extract obtained in (A) to acidic pH, heating the crude extract and then filtering or centrifuging the crude extract to conduct a deproteinizing treatment, after that, adjusting the deproteinized solution to basic pH, heating the deproteinized solution and then filtering or centrifuging the deproteinized solution to give a deproteinized filtrate or supernatant,
• (C) adjusting the filtrate or the supernatant obtained in (B) to pH 3.5 to 5.5 and adsorbing the filtrate or the supernatant with an adsorbent, and
• (D) adding an extraction solvent to the adsorbent obtained in (C) to obtain a mixture, adjusting the mixture to pH 9 to 12, and eluting the adsorbed component to give the extract.

A method for providing an analgesic effect comprising administering a preparation containing an extract from inflamed skins of rabbits inoculated with vaccinia virus, in which the variations in the content of N-acetylneuraminic acid contained in a preparation among manufacturing lots are reduced by an amount of N-acetylneuraminic acid contained in the preparation being measured for each manufacturing lot and the content is confirmed to be 4000 ng or more per unit of the extract in the preparation, to a subject in need thereof,

• wherein the extract is obtained by a manufacturing method comprising:
• (A) collecting inflamed skin tissues of rabbits subjected to the intradermal inoculation with vaccinia virus, and crushing the inflamed tissues, conducting an extraction treatment comprising adding to the crushed tissues an extraction solvent in 1- to 5-fold thereof by volume of water, phenol water, physiological saline or phenol-added glycerin water to obtain a mixture, allowing the mixture to stand for 3 to 7 days and further heating the mixture at 35 to 40° C., then, filtrating or centrifuging the mixture to give a crude extract which is a filtrate or supernatant, and removing tissue fragments from the crude extract,
• (B) adjusting the crude extract obtained in (A) to acidic pH, heating the crude extract and then filtering or centrifuging the crude extract to conduct a deproteinizing treatment, after that, adjusting the deproteinized solution to basic pH, heating the deproteinized solution and then filtering or centrifuging the deproteinized solution to give a deproteinized filtrate or supernatant,
• (C) adjusting the filtrate or the supernatant obtained in (B) to pH 3.5 to 5.5 and adsorbing the filtrate or the supernatant with an adsorbent, and
• (D) adding an extraction solvent to the adsorbent obtained in (C) to obtain a mixture, adjusting the mixture to pH 9 to 12, and eluting the adsorbed component to give the extract.

A method for controlling the manufacture of a preparation containing an extract from inflamed skins of rabbits inoculated with vaccinia virus in which the variations in the content of N-acetylneuraminic acid contained in the preparation among manufacturing lots are reduced, comprising measuring the amount of N-acetylneuraminic acid contained in the preparation for each manufacturing lot, and only qualifying preparations having a N-acetylneuraminic acid content which is 4000 ng or more per unit of the extract in the preparation,

• wherein the extract is obtained by a manufacturing method comprising:
• (A) collecting inflamed skin tissues of rabbits subjected to the intradermal inoculation with vaccinia virus, and crushing the inflamed tissues, conducting an extraction treatment comprising adding to the crushed tissues an extraction solvent in 1- to 5-fold thereof by volume of water, phenol water, physiological saline or phenol-added glycerin water to obtain a mixture, allowing the mixture to stand for 3 to 7 days and further heating the mixture at 35 to 40° C., then, filtrating or centrifuging the mixture to give a crude extract which is a filtrate or supernatant, and removing tissue fragments from the crude extract,
• (B) adjusting the crude extract obtained in (A) to acidic pH, heating the crude extract and then filtering or centrifuging the crude extract to conduct a deproteinizing treatment, after that, adjusting the deproteinized solution to basic pH, heating the deproteinized solution and then filtering or centrifuging the deproteinized solution to give a deproteinized filtrate or supernatant,
• (C) adjusting the filtrate or the supernatant obtained in (B) to pH 3.5 to 5.5 and adsorbing the filtrate or the supernatant with an adsorbent, and
• (D) adding an extraction solvent to the adsorbent obtained in (C) to obtain a mixture, adjusting the mixture to pH 9 to 12, and eluting the adsorbed component to give the extract.