There are important points when preparing liposome formulations.
Uniformity at the molecular level is required. [Preventing cholesterol precipitation]
The organic solvent must be completely removed.
It must be fully hydrated.
In the lab, liposomes can be forcibly prepared using a variety of methods and equipment, but in most cases, scale-up production cannot be performed as in the lab.
Our mixed lipid "Presome™" eliminates many of these problems.
About Presome
Presome is a lipid premix that can greatly reduce the complexity of liposome production. Simply add an aqueous solution without using an organic solvent and stir gently to obtain an MLV liposome solution with a particle size of about 1㎛.
After this, the desired liposomes can be obtained by sizing to the desired particle size as necessary.
Presome is not just a lipid powder mixture or a solvent mixture from a lipid mixture solution, but a premix made through a special process for preparing liposomes. And Presome is ideal for simplifying the liposome production process, mass production, and preparation of highly concentrated solutions.
Merit of using Presome™
An example of liposome preparation on a small scale
Method
Overview
Merit / Demerit
Bangham method
Dissolve lipid in organic solvent such as chloroform,
Hydrated by forming a thin film in Eggplant flask with an evaporator.
○Relatively easy in the lab
×Use organic solvent [remains in liposome solution]
×Sonication required
×Difficult to scale up
×Risk of cholesterol deposition during thin film formation
Reverse phase evaporation method
Dissolve lipid in organic solvent such as chloroform,
Slowly add aqueous solution to this and use ultrasonic waves etc.
Disperse and recover the solvent at reduced pressure.
×Use organic solvent [remains in liposome solution]
×Sonication required
Mechanochemical method
Refine by shearing force using an ultra-high pressure homogenizer.
○Need not organic solvent
×Pretreatment is required to make lipid and aqueous solution compatible
×Requires many processing times
×Large scale equipment is required for scale-up
Since an organic solvent is used when preparing liposomes, there are many problems such as solvent remaining in the liposome solution, precipitation of cholesterol, difficulty in scaling up, and large equipment.
Use Presome™ to solve these problems!!
The complicated operation of liposome solution production can be saved greatly.
Liposome liquid mass production and high concentration liquid preparation are possible.
Production of up to 20㎏ / lot is possible.
Presome can be manufactured with the desired lipid composition.
Presome containing fatty-soluble APIs may also be adjustable.
Presome is an original technology of NFC.
Material lipids such as NFC's high purity phospholipids, high purity cholesterol, and high purity PEG lipids can be used.
Consistent quality control / GMP management is possible.
Two samples were hydrated by shaking in a 65℃ bath.
Shaking continued until the powder was completely melted.
Both were shaken at 65℃ for 6 minutes.
The mixed product was very foamy, and the Presome method had almost no foaming.
Lipid content = 0.5g / 13.5mL [250mM Ammonium sulfate aqueous] solution
Two samples were sized using an extruder heated at 65℃.
The entire mixture passed in one minute, but the Presome method took about 3 minutes.
Using filter type: 50㎚ polycarbonate filter
Liquid volume: Approximately 14mL [use 0.5g of total lipid weight]
Using filter type: 50㎚ polycarbonate filter
Liquid volume: Approximately 14mL [use 0.5g of total lipid weight]
The result of sizing … There was nothing in the Presome method
When the extruder was disassembled after sizing, many crystals remained on the filter in the mixed product, but there was nothing in the Presome method.
Is the mixed product not fully hydrated?
The lipid materials used were not partially hydrated and existed as crystals [solids], and were filtered out without passing through the filter when sizing with the extruder. Visually, about half of the lipids used were filtered out.
In general, cholesterol is highly fatty-soluble, and will not dissolve sufficiently and will not be incorporated into lipid membranes by simply mixing it with water or mixing it with other phospholipids. In order to confirm this, We measured the lipid composition analysis by HPLC and IR to know the difference in chemical structure.
※HSPC is a hydrogenated soybean phospholipid, and it is a mixture of DSPC and PSPC.Enlarge illustration
Only lipid mixture will change lipid composition
The lipid composition of the liposome solution obtained by HPLC / UV detector was confirmed semi-quantitatively.
HPLC conditions
Mobile phase: reverse phase condition
Detector: UV [215㎚]
Column: Reverse phase column
Temperature: 40℃
Flow rate: 0.6mL
As a result, almost no peak derived from cholesterol [RT = 3.4min] was confirmed in the mixed product, whereas the peak derived from cholesterol [RT = 3.4min] and MPEG2000-DSPE [RT = 1.7min] in the Presome method.
And two peaks [RT = 3.9 and 4.9min] derived from HSPC were confirmed by the specified amount.
After sizing with the mixture, what remained on the extruder filter was cholesterol.
If each lipid was simply mixed and hydrated, cholesterol crystals were mainly filtered during sizing and remained on the filter as crystals. As a result, the liposome composition has changed.
It is confirmed that the liposome solution obtained by hydration and sizing using PresomeACD-1 does not remain on the filter, and cholesterol is taken into the lipid membrane and is normally liposome-formed. it is complete.
※HSPC is a hydrogenated soybean phospholipid, and it is a mixture of DSPC and PSPC.
The chemical structure changes when it becomes Presome
When the IR of Presome ACD-1 was measured with a simple mixture of lipids, it was found that there were some differences between the two.
Changes appear mainly in the fingerprint region near 1300 - 600㎝-1, which is considered to be a change in crystal structure, not a change in functional groups.
The differences in IR charts suggest that each component of the lipid is not a mixture of powders with a single crystal structure, but the crystalline bonds of each component are lost and new intermolecular bonds are formed.
Summary of Presome™ Hydration Capability
It was found that Presome is not just a mixture of lipid components, but an easily hydrated solid that is mixed to the molecular level.
Presome does not require any special pretreatment and can be hydrated by mixing with aqueous solution and stirring.
An example of Presome™
Product name
Lipid Content
Presome ACD-1
HSPC: Cholesterol: MPEG-DSPE
Presome PPG-Ⅰ
DPPC: Cholesterol: DPPG
Presome SST-Ⅱ
DSPC: Cholesterol: MPEG-DSPE
Presome MMG-Ⅱ
DMPC: Cholesterol: DMPG
Presome EEG-Ⅰ
PCE: Cholesterol: PGE
Presome TSU
DPPC: Cholesterol
The above composition is an example. In addition to these, it is possible to manufacture with the desired lipid composition.
Presome including substances such as fat-soluble API may be prepared.
Presome ACD-1 is sold through Tokyo Chemical Industry Co., Ltd as a reagent. If you are interested in this compound, please visit the following address.
When you request its GMP-grade product, please access by contact form shown in the bottom of this page.
Presome™ manufacturing equipment
In the production of Presome used for pharmaceuticals, all operations are performed in the environmental management area. After the raw material is dissolved in a solvent, the product obtained by processing with the CRUX system is dried in a dryer and packaged.
The process of manufacturing Presome is approved by FDA and EMA.
Presome manufacturing room [environmental controlled area]Presome manufacturing equipment "CRUX system"Presome product outletShed-type vacuum dryer [capacity of about 50㎏]
Contact
For inquiries about Presome, please contact us using the form below.
![Presome manufacturing room [environmental controlled area]](https://www.nipponseika.co.jp/en/business/healthcare/images/presome/img_09.jpg)
![Shed-type vacuum dryer [capacity of about 50㎏]](https://www.nipponseika.co.jp/en/business/healthcare/images/presome/img_12.jpg)