A pharmaceutical formulation and preparation process thereof

Abstract

The invention relates to a drug formed by 106-1013vp / ml virus particles and findings, wherein the findings comprise microcosmic salt buffer system, 10-200mM NaCl, 1-2mM MgCl2 or CaCl2, 10-500 muM EDTA, 0.15-1% absolute ethyl alcohol, 0.01-0.09% Tween 80, 0-21mMhistidine, and glycerin, while it can also contain L-aminothiopropionic acid. The invention can be stored more than 3 years under 8Deg. C.

Description

technical field

[0001] The invention belongs to the field of pharmaceutical preparations and preparations, and in particular relates to a recombinant virus preparation and a preparation method thereof. Background technique

[0002] With the launch of Jinyousheng, the world's first gene therapy product, in 2004, gene therapy has been increasingly recognized by people. Gene therapy has three components: the target gene, the carrier that carries the gene into the cell for expression, and the target cell. Among them, the lack of specific, targeted and efficient gene transfer vectors has always been a difficult problem in gene therapy. At present, the vectors used in gene therapy research are mainly divided into two categories: viral vectors and non-viral vectors. Non-viral vectors mainly refer to liposomes and naked DNA. The development of the gene transduction system has been paid attention to in recent years due to the characteristics of unrestricted transferable genes, safe...

Examples

Embodiment 1

[0030] Embodiment 1: Preparation formula selection experiment

[0031] Recombinant human hepatitis B core antigen adenovirus preparation prescription:

[0032] Prescription 1: KH 2 PO 4 0.41g; K 2 HPO 4 5.59g; NaCl 4.38g;

[0033] MgCl 2 ·6H 2 O 0.20g; Glycerin 100ml; Tween80 0.2ml;

[0034] EDTA 0.03g; ethanol 5ml; pH7.5

[0035] Prescription 2: KH 2 PO 4 0.41g; K 2 HPO 4 5.59g; NaCl 9.00g;

[0036] Histidine 1.55g; Glycerin: 100ml; pH7.5

[0037] Prescription 3: KH 2 PO 4 0.41g; K 2 HPO 4 5.59g; NaCl 4.38gMgCl 2 ·6H 2 O: 0.20g; pH7.5

[0038] Prescription 4: KH2PO4 0.41g; K2HPO4 5.59g; NaCl 9.00g;

[0039] Glycerin 100ml; Tween80: 0.2ml; L-cysteine ​​1.20g; pH7.5 Preparation:

[0040] Weigh each component according to the formula, add distilled water to about 500ml, stir well, wait until it is completely dissolved, and then make the purified concentration 5×10 11 Add 10ml of vp / ml recombinant human hepatitis B core antigen adenovirus preparation ...

Embodiment 2

[0044] Embodiment 2: formulation accelerated test (temperature-time test)

[0045] By accelerating the chemical or physical changes of the drug, the stability of the drug is predicted.

[0046] A certain amount of prescription 1 preparations were taken from the same batch of preparations and stored under different temperature conditions: -80°C, -20°C, 8°C, 37°C. The preparations are marked with the batch number and storage temperature. On the day of storage at different temperatures (week 0), samples were taken for testing, and then samples were taken for TCID50 testing according to the time points in the following table.

[0047] Table 1: Detection frequency of different temperature conditions in the accelerated test of preparations

[0048] temperature

[0049] The test results are shown in Figure 4. It can be seen from the figure that the quality of the preparation is stable at -80°C, -20°C and 8°C for 24 weeks, and the test results fluctuate within the measur...

Embodiment 3

[0050] Embodiment 3: long-term stability test

[0051] The long-term test is carried out under the conditions close to the actual storage of the drug, and its purpose is to provide a basis for formulating the effective period of the drug.

[0052] Based on the results of Example 3, this experiment was designed: 8°C was selected for long-term stability experiment. Store three batches of prescription 1 preparations at 8°C, take samples for testing on the day of storage (0 month), and then take samples for TCID50 testing at 1, 3, 6, 9, 12, 18, 24, and 36 months of storage . The test results are shown in Table 2:

[0053] Table 2: Long-term stability test results stored at 8°C: (unit: IU / ml)

[0054] time

[0055] According to the experimental results, see Figure 5. The results show that: the adenovirus liquid preparation made by prescription 1 of the present invention has good stability, and it is stored for 36 months (i.e. 3 years) at 8°C, and the infectivity tite...

technical field

[0001] The invention belongs to the field of pharmaceutical preparations and preparations, and in particular relates to a recombinant virus preparation and a preparation method thereof. Background technique

[0002] With the launch of Jinyousheng, the world's first gene therapy product, in 2004, gene therapy has been increasingly recognized by people. Gene therapy has three components: the target gene, the carrier that carries the gene into the cell for expression, and the target cell. Among them, the lack of specific, targeted and efficient gene transfer vectors has always been a difficult problem in gene therapy. At present, the vectors used in gene therapy research are mainly divided into two categories: viral vectors and non-viral vectors. Non-viral vectors mainly refer to liposomes and naked DNA. The development of the gene transduction system has been paid attention to in recent years due to the characteristics of unrestricted transferable genes, safe...

Examples

Embodiment 1

[0030] Embodiment 1: Preparation formula selection experiment

[0031] Recombinant human hepatitis B core antigen adenovirus preparation prescription:

[0032] Prescription 1: KH 2 PO 4 0.41g; K 2 HPO 4 5.59g; NaCl 4.38g;

[0033] MgCl 2 ·6H 2 O 0.20g; Glycerin 100ml; Tween80 0.2ml;

[0034] EDTA 0.03g; ethanol 5ml; pH7.5

[0035] Prescription 2: KH 2 PO 4 0.41g; K 2 HPO 4 5.59g; NaCl 9.00g;

[0036] Histidine 1.55g; Glycerin: 100ml; pH7.5

[0037] Prescription 3: KH 2 PO 4 0.41g; K 2 HPO 4 5.59g; NaCl 4.38gMgCl 2 ·6H 2 O: 0.20g; pH7.5

[0038] Prescription 4: KH2PO4 0.41g; K2HPO4 5.59g; NaCl 9.00g;

[0039] Glycerin 100ml; Tween80: 0.2ml; L-cysteine ​​1.20g; pH7.5 Preparation:

[0040] Weigh each component according to the formula, add distilled water to about 500ml, stir well, wait until it is completely dissolved, and then make the purified concentration 5×10 11 Add 10ml of vp / ml recombinant human hepatitis B core antigen adenovirus preparation ...

Embodiment 2

[0044] Embodiment 2: formulation accelerated test (temperature-time test)

[0045] By accelerating the chemical or physical changes of the drug, the stability of the drug is predicted.

[0046] A certain amount of prescription 1 preparations were taken from the same batch of preparations and stored under different temperature conditions: -80°C, -20°C, 8°C, 37°C. The preparations are marked with the batch number and storage temperature. On the day of storage at different temperatures (week 0), samples were taken for testing, and then samples were taken for TCID50 testing according to the time points in the following table.

[0047] Table 1: Detection frequency of different temperature conditions in the accelerated test of preparations

[0048] temperature

[0049] The test results are shown in Figure 4. It can be seen from the figure that the quality of the preparation is stable at -80°C, -20°C and 8°C for 24 weeks, and the test results fluctuate within the measur...

Embodiment 3

[0050] Embodiment 3: long-term stability test

[0051] The long-term test is carried out under the conditions close to the actual storage of the drug, and its purpose is to provide a basis for formulating the effective period of the drug.

[0052] Based on the results of Example 3, this experiment was designed: 8°C was selected for long-term stability experiment. Store three batches of prescription 1 preparations at 8°C, take samples for testing on the day of storage (0 month), and then take samples for TCID50 testing at 1, 3, 6, 9, 12, 18, 24, and 36 months of storage . The test results are shown in Table 2:

[0053] Table 2: Long-term stability test results stored at 8°C: (unit: IU / ml)

[0054] time

[0055] According to the experimental results, see Figure 5. The results show that: the adenovirus liquid preparation made by prescription 1 of the present invention has good stability, and it is stored for 36 months (i.e. 3 years) at 8°C, and the infectivity tite...

Images