[go: up one dir, main page]

WO2020037752A1 - Vert d'indocyanine modifié et son procédé de préparation et son utilisation - Google Patents

Vert d'indocyanine modifié et son procédé de préparation et son utilisation Download PDF

Info

Publication number
WO2020037752A1
WO2020037752A1 PCT/CN2018/106522 CN2018106522W WO2020037752A1 WO 2020037752 A1 WO2020037752 A1 WO 2020037752A1 CN 2018106522 W CN2018106522 W CN 2018106522W WO 2020037752 A1 WO2020037752 A1 WO 2020037752A1
Authority
WO
WIPO (PCT)
Prior art keywords
icg
mpeg
modified
peg
indocyanine green
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Ceased
Application number
PCT/CN2018/106522
Other languages
English (en)
Chinese (zh)
Inventor
程震
娄宏跃
刘弘光
索永宽
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Northeastern University China
Original Assignee
Northeastern University China
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Northeastern University China filed Critical Northeastern University China
Publication of WO2020037752A1 publication Critical patent/WO2020037752A1/fr
Anticipated expiration legal-status Critical
Ceased legal-status Critical Current

Links

Images

Classifications

    • C—CHEMISTRY; METALLURGY
    • C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
    • C08G—MACROMOLECULAR COMPOUNDS OBTAINED OTHERWISE THAN BY REACTIONS ONLY INVOLVING UNSATURATED CARBON-TO-CARBON BONDS
    • C08G65/00—Macromolecular compounds obtained by reactions forming an ether link in the main chain of the macromolecule
    • C08G65/02—Macromolecular compounds obtained by reactions forming an ether link in the main chain of the macromolecule from cyclic ethers by opening of the heterocyclic ring
    • C08G65/32—Polymers modified by chemical after-treatment
    • C08G65/329—Polymers modified by chemical after-treatment with organic compounds
    • C08G65/334—Polymers modified by chemical after-treatment with organic compounds containing sulfur
    • C08G65/3348—Polymers modified by chemical after-treatment with organic compounds containing sulfur containing nitrogen in addition to sulfur
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K49/00—Preparations for testing in vivo
    • A61K49/001—Preparation for luminescence or biological staining
    • A61K49/0013—Luminescence
    • A61K49/0017—Fluorescence in vivo
    • A61K49/0019—Fluorescence in vivo characterised by the fluorescent group, e.g. oligomeric, polymeric or dendritic molecules
    • A61K49/0021—Fluorescence in vivo characterised by the fluorescent group, e.g. oligomeric, polymeric or dendritic molecules the fluorescent group being a small organic molecule
    • A61K49/0032—Methine dyes, e.g. cyanine dyes
    • A61K49/0034—Indocyanine green, i.e. ICG, cardiogreen
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K49/00—Preparations for testing in vivo
    • A61K49/001—Preparation for luminescence or biological staining
    • A61K49/0013—Luminescence
    • A61K49/0017—Fluorescence in vivo
    • A61K49/005—Fluorescence in vivo characterised by the carrier molecule carrying the fluorescent agent
    • A61K49/0054—Macromolecular compounds, i.e. oligomers, polymers, dendrimers
    • C—CHEMISTRY; METALLURGY
    • C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
    • C08G—MACROMOLECULAR COMPOUNDS OBTAINED OTHERWISE THAN BY REACTIONS ONLY INVOLVING UNSATURATED CARBON-TO-CARBON BONDS
    • C08G2650/00—Macromolecular compounds obtained by reactions forming an ether link in the main chain of the macromolecule
    • C08G2650/02—Macromolecular compounds obtained by reactions forming an ether link in the main chain of the macromolecule characterized by the type of post-polymerisation functionalisation
    • C08G2650/04—End-capping
    • C—CHEMISTRY; METALLURGY
    • C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
    • C08G—MACROMOLECULAR COMPOUNDS OBTAINED OTHERWISE THAN BY REACTIONS ONLY INVOLVING UNSATURATED CARBON-TO-CARBON BONDS
    • C08G2650/00—Macromolecular compounds obtained by reactions forming an ether link in the main chain of the macromolecule
    • C08G2650/28—Macromolecular compounds obtained by reactions forming an ether link in the main chain of the macromolecule characterised by the polymer type
    • C08G2650/50—Macromolecular compounds obtained by reactions forming an ether link in the main chain of the macromolecule characterised by the polymer type containing nitrogen, e.g. polyetheramines or Jeffamines(r)

Definitions

  • the invention relates to a modification method of vascular fluorescence contrast agent, in particular to indigocyanine green modified with PEG, and a preparation method and application thereof.
  • Indocyanine Green is a tricarbonyl cyanine dye with a molecular weight of 751.4 Da. It is a negatively charged ion and belongs to the family of cyanine dyes.
  • the indocyanine green structural formula is shown in FIG. 1A of the accompanying drawings. It is a dye medicine commonly used to check liver function and liver effective blood flow. Immediately after being injected into the body, it is combined with plasma proteins, and is quickly distributed in the blood vessels of the whole body with blood circulation. It is efficiently and selectively taken up by hepatocytes, and then excreted from the hepatocytes into the bile in a free form, enters the intestine through the biliary tract, and is discharged with feces. in vitro.
  • ICG is widely used in surgery for angiography. Compared with other angiography methods (X-ray, CT, MRI and PET), ICG can be easily and economically injected for angiography, such as neurosurgery, bypass coronary surgery , Flap surgery, reconstruction surgery, trauma surgery and laparoscopic surgery, used to check blood circulation and intraoperative navigation.
  • the ICG bolus enters the systemic blood circulation, and imaging is completed within minutes after the injection, and it can usually be re-injected after about 15 minutes as needed.
  • ICG is also used in tumor detection and treatment.
  • ICG's fluorescence quantum yield is about 10%, and the fluorescence and photoacoustic signals increase and decrease with the fluorescence quantum yield, respectively.
  • ICG can be used as a contrast agent for fluorescence imaging and photoacoustic imaging.
  • ICG can use the EPR effect to non-specifically aggregate in tumors, or target modification, specifically to aggregate in tumors, so as to perform tumor detection, localization, and photothermal therapy and photodynamic therapy.
  • the ICG has a very short half-life in plasma, which limits the application of ICG as a near-infrared fluorescent imaging material.
  • clinical practice is based on the use of multiple injections or slow injections to obtain accurate fluorescent detection results for the clinic. difficult.
  • 25 mg of ICG is dissolved in as little sterilized injection water as possible, diluted with physiological saline to a concentration of 2.5 to 5.0 mg / ml, and the above-mentioned solution equivalent to 3 mg of ICG is injected intravenously.
  • the intravenous infusion was continued at a rate of 0.27 to 0.49 mg per minute for about 50 minutes at a certain rate until the blood sample was taken (both peripheral blood and hepatic vein blood were required).
  • the present invention is expected to modify or modify the existing ICG to extend the plasma half-life, eliminating the need to give patients multiple injections or slow and continuous injections in the blood fluoroscopy test; at the same time, the modified or The modified ICG has good fluorescence properties and spectral absorption capabilities close to or better than the original ICG, and does not affect its application characteristics in angiography.
  • a modified indocyanine green in which one of the two sulfonic groups in the molecular structure of the indocyanine green is substituted with a modified PEG group.
  • the modified PEG is also called modified polyethylene glycol, which is a PEG modified by a chemical modification group or a biologically active group.
  • the modified PEG group refers to an active group formed after the modified PEG loses an H atom.
  • the modified PEG includes, but is not limited to, mPEG (monomethoxy ether PEG), mPEG-NH 2 (mPEG-amino), mPEG-SS (mPEG-succinimide) Succinate), mPEG-SC (mPEG-succinimide carbonate), mPEG2-NHS (mPEG2-N-hydroxysuccinimide ester), mPEG-SPA (mPEG-succinimide propionate) , MPEG-ALD (mPEG-propionaldehyde), mPEG-MAL (mPEG-maleimide), HO-PEG-COOH (a-hydroxy-w-carboxy polyethylene glycol), mPEG-b-PS (poly Block copolymer of ethylene glycol and polystyrene), mPEG-b-PI (block copolymer of polyethylene glycol and polyisoprene), mPEG-b-
  • the molecular weight of the modified PEG group ranges from 500 to 20,000, preferably 500, 2000, 5000, or 10,000, 20,000.
  • the molecular structural formula of the PEG-modified indocyanine green is represented as follows:
  • n 9 to 450.
  • a method for preparing a modified indocyanine green is proposed.
  • the molecular structure formula of the modified indocyanine green is as follows:
  • ICG derivative modified with -CO-NHS ester group and mPEG-NH 2 are used as reactants, in which mPEG-NH 2 undergoes an ester substitution reaction with -CO-NHS in the ICG derivative to remove a molecule of N-hydroxyl Succinimide, a new type of ICG modified by mPEG-NH- (see structural formula C); the reaction process is expressed as:
  • the reaction conditions in the above reaction process are: the -CO-NHS ester group-modified ICG derivative is mixed with mPEG-NH 2 at a molar ratio of 1: 5, at a pH of 7.0 to In 8.0 PBS, the reaction was stirred at room temperature for 5-12 hours, and purified by HPLC and other methods to obtain PEG-ICG (see structural formula C).
  • the main invention of the present invention is to replace one of the two sulfonic acid groups of the indocyanine green molecular structure with a modified PEG group to extend the plasma half-life of ICG without affecting or even strengthening it.
  • the sulfonic acid group of ICG is difficult to directly replace with modified PEG. Therefore, the preparation method of the present invention uses the ICG derivative modified with -CO-NHS ester group as the reactant instead of ICG.
  • the reaction is highly controllable, and the reaction product can be designed according to needs, and the reaction conditions are mild (room temperature).
  • the synthetic route has high selectivity and yield, and can be easily converted for industrial production.
  • one of the sulfonic acid groups -SO 3 H of ICG is substituted with a modified PEG group, thereby achieving the effect of significantly extending the half-life of ICG in human plasma.
  • PEG-ICGs and ICG were injected into the tail vein of mice, and near-infrared region II fluorescence imaging was performed on the femoral veins of the mice. From 0.06421h of unmodified ICG to 1.12731h to 6.2413h, compared to unmodified ICG, ICG modified with modified PEG groups with molecular weights of 500, 2000, 5000, 10,000, and 20,000 has a longer retention time in blood. Significantly extended, optimizing its applications in angiography, intraoperative navigation, tumor detection and treatment.
  • the modified and unmodified ICGs have basically the same spectral absorption capabilities, and are in the near infrared I region (wavelength 650nm-900nm) and II region (wavelength 900nm). -1200nm) have good fluorescence properties, and are consistent with the absorption and emission spectrum properties of unmodified ICG. This shows that PEG-ICGs have the same application value of near-infrared I and II region imaging as ICG.
  • modified PEG-ICGs showed good stability in both PBS (phosphate buffered saline) and FBS (fetal bovine serum) at pH 7.4, so the modified PEG-ICGs of the present invention have Prospects for commercial production and applications.
  • PBS phosphate buffered saline
  • FBS fetal bovine serum
  • FIG. 1A is a molecular structural formula of indocyanine green ICG
  • FIG. 1B is a molecular structural formula of modified PEG-ICGs according to the present invention.
  • FIG. 2A shows that the intermediate product ICG-NHS of modified ICG is purified by high-performance liquid chromatography analysis and an absorption peak is detected in a 780 nm channel.
  • Figure 2B, Figure 2C, Figure 2D, Figure 2E, and Figure 2F are PEG500-ICG, PEG2000-ICG, PEG5000-ICG, and PEG10000-ICG, respectively, obtained by mPEG-NH 2 with molecular weights of 500, 2000, 5000, 10,000, and 20,000.
  • ICG and PEG20000-ICG were purified by HPLC analysis and the absorption peaks were detected in the 780nm channel.
  • Figure 3 shows the relative absorption spectra of unmodified ICG and modified PEG500-ICG, PEG2000-ICG, PEG5000-ICG, PEG10000-ICG, and PEG20000-ICG at a concentration of 20 ⁇ g / ml at a wavelength of 600 nm to 950 nm.
  • Figure 4 shows the relative fluorescence intensities of unmodified ICG and modified PEG500-ICG, PEG2000-ICG, PEG5000-ICG, PEG10000-ICG, and PEG20000-ICG in the near-infrared I region (wavelength 650nm-900nm).
  • Figure 5 shows the relative fluorescence intensities of unmodified ICG and modified PEG500-ICG, PEG2000-ICG, PEG5000-ICG, PEG10000-ICG, and PEG20000-ICG in the near infrared II region (wavelength 900nm-1200nm).
  • Fig. 6 shows the decrease rate of fluorescence intensity of modified PEG500-ICG, PEG2000-ICG, PEG5000-ICG, PEG10000-ICG, and PEG20000-ICG in PBS (components Na 2 HPO 4 and NaH 2 PO 4 ) at pH 7.4. Time variation curve.
  • FIG. 7 is a graph showing the change rate of the fluorescence intensity decrease rate of FPEG (fetal bovine serum) with time for modified PEG500-ICG, PEG2000-ICG, PEG5000-ICG, PEG10000-ICG, and PEG20000-ICG.
  • FPEG fetal bovine serum
  • Figure 8 shows unmodified ICG and modified PEG500-ICG, PEG2000-ICG, PEG5000-ICG, PEG10000-ICG, and PEG20000-ICG in the near-infrared region II after mice were injected with 2 mg ⁇ mL -1 tail vein. Group of fluorescence imaging photos.
  • Fig. 9 is a photo of fluorescence imaging of near-infrared region II in mice with tumors by injecting unmodified ICG and modified PEG5000-ICG into the tail vein, respectively, and testing the EPR of modified PEG5000-ICG. effect.
  • the vascular fluorescent imaging material ICG is mainly modified with a modified PEG group to extend its half-life in the blood.
  • Modified PEG is also called modified polyethylene glycol. It is a PEG modified by a chemical modification group or a biologically active group, that is, the conventional PEG changes its terminal group, and -OH, -COOH, -NH 2 , methoxy, nitrile and the like are substituted or linked.
  • the modified PEG group refers to an active group formed after the modified PEG loses the H atom at the terminal.
  • Modified PEGs include, but are not limited to, mPEG (monomethoxy ether PEG), mPEG-NH 2 (mPEG-amine), mPEG-SS (mPEG-succinimide succinate), mPEG-SC (mPEG-amber Imide carbonate), mPEG2-NHS (mPEG2-N-hydroxysuccinimide ester), mPEG-SPA (mPEG-succinimide propionate), mPEG-ALD (mPEG-propionaldehyde), mPEG -MAL (mPEG-maleimide), HO-PEG-COOH (a-hydroxy-w-carboxy polyethylene glycol), mPEG-b-PS (block copolymer of polyethylene glycol and polystyrene) , MPEG-b-PI (block copolymer of polyethylene glycol and polyisoprene), mPEG-b-PAN (block copolymer of polyethylene glycol
  • the molecular structure formula of the modified indocyanine green of the present invention is represented as follows:
  • n 9 to 450.
  • the method for preparing the above-mentioned PEG-modified indocyanine green using the ICG derivative and mPEG-NH 2 modified by the -CO-NHS ester group as reactants, wherein mPEG-NH 2 and the ICG derivative are reacted -CO-NHS undergoes an ester substitution reaction to remove a molecule of N-hydroxysuccinimide to prepare a new type of ICG modified by mPEG-NH- (see structural formula C); the reaction process is expressed as:
  • the ICG (see structural formula C) modified by the modified PEG group of the present invention is significantly prolonged in the blood, which optimizes its applications in angiography, intraoperative navigation, tumor detection and treatment, and other aspects.
  • This example is the same as Preparation Example 1, except that mPEG-NH 2 with a molecular weight of about 2000 was used for the reaction.
  • the product is designated as PEG2000-ICG.
  • This example is the same as Preparation Example 1, except that mPEG-NH 2 with a molecular weight of about 5000 was used for the reaction.
  • the product is designated as PEG5000-ICG.
  • This example is the same as Preparation Example 1, except that mPEG-NH 2 with a molecular weight of about 10,000 was used for the reaction.
  • the product is designated as PEG10000-ICG.
  • This example is the same as Preparation Example 1, except that mPEG-NH 2 with a molecular weight of about 20,000 is used for the reaction.
  • the product is designated as PEG20000-ICG.
  • the PEG500-ICG, PEG2000-ICG, PEG5000-ICG, PEG10000-ICG and PEG20000-ICG prepared in Preparation Examples 1 to 5 are purified by high performance liquid chromatography (HPLC), and a separation column (C4 Column, 300A, 3.5 ⁇ m, 4.6 mm ⁇ 250 mm), and the absorption peak was detected under the 780 nm channel.
  • HPLC high performance liquid chromatography
  • C4 Column 300A, 3.5 ⁇ m, 4.6 mm ⁇ 250 mm
  • Figure 2A is an HPLC chart of the reactant ICG-NHS (ICG modified with -CO-NHS ester group) of structural formula B.
  • the retention time in the separation column is 22.4943min.
  • ICG-NHS has low polarity and lipophilicity. Poor water solubility and other characteristics.
  • FIG. 2B is an HPLC chart of PEG500-ICG in Preparation Example 1.
  • the peak time is 20.7236min earlier than ICG-NHS, which indicates that the polarity of ICG is relatively larger and the water solubility is better, which proves that mPEG-NH 2 and The ICG connection was successful. Since mPEG-NH 2 are not connected to the ICG is no specific absorption peak, it is not detected by HPLC mPEG-NH 2.
  • the peak time of Figure 2B is earlier than ICG-NHS, which proves that mPEG-NH 2 is successfully connected to ICG.
  • FIG. 2C is an HPLC chart of PEG2000-ICG in Preparation Example 2.
  • the peak time is 19.6685min earlier than ICG-NHS and PEG500-ICG. It also shows that mPEG-NH 2 has a larger molecular weight, which further polarizes ICG and changes its water solubility. Good, so the peak time of PEG2000-ICG is earlier than that of PEG500-ICG, which proves that mPEG-NH 2 is successfully connected to ICG.
  • FIG. 2D is an HPLC chart of the PEG5000-ICG chart of Preparation Example 3.
  • the peak time was 19.2725min earlier than ICG-NHS, PEG500-ICG, and PEG2000-ICG.
  • mPEG-NH 2 was successfully connected to ICG.
  • FIG. 2E is an HPLC chart of PEG10000-ICG in Preparation Example 4.
  • the peak time is 19.6829min earlier than ICG-NHS, but the peak time is later than PEG5000-ICG. It may be because the molecular weight of the connected mPEG-NH 2 is too large. Due to the position in the column, although the water solubility is better, the peak time is later than PEG5K-ICG. But again, mPEG-NH 2 that has not been successfully connected to ICG has no specific absorption peak, so FIG. 2E can prove that mPEG-NH 2 is successfully connected to ICG.
  • FIG. 2F is an HPLC chart of PEG20000-ICG in Preparation Example 5.
  • the peak time is 19.4424min.
  • the peak time is earlier than ICG-NHS, and the peak time is later than PEG5K-ICG. It may also be because the molecular weight of the attached PEG is too large, so although the water solubility is better, the peak time is later than PEG5K-ICG. But for the same reason, FIG. 2F can prove that mPEG-NH 2 is successfully connected with ICG.
  • PEG-ICGs can be detected only after the connection is successful, and the peak time is earlier than that of ICG-NHS, indicating that the connection is successful, and the PEG-ICGs after the successful connection have better water solubility than ICG-HNS. .
  • PEG500-ICG, PEG2000-ICG, PEG5000-ICG, PEG10000-ICG, and PEG20000-ICG prepared in Preparation Example 1-5 were tested for their spectral properties (including absorption spectral properties and emission spectral properties), and compared with unmodified ICG Compare.
  • the specific operation is as follows:
  • the UV-Vis-NIR spectrometer cary 5000 was used to measure the spectral absorption intensities of ICG, PEG500-ICG, PEG2000-ICG, PEG5000-ICG, PEG10000-ICG, and PEG20000-ICG in the spectral range of 500nm-1000nm, respectively. See Figure 3.
  • the weighted continuous curve represents PEG20000-ICG
  • the gray continuous line represents ICG
  • the dark heavy dotted line represents PEG500-ICG
  • the thin dotted line represents PEG2000-ICG
  • the one-dot chain line represents PEG5000-ICG
  • the two-dot chain line represents PEG10000-ICG.
  • the modified PEG-ICGs exhibited a spectral absorption ability substantially consistent with that of the unmodified ICG.
  • the spectrophotometer 200 set the excitation light at 808nm, and test the relative of ICG, PEG500-ICG, PEG2000-ICG, PEG5000-ICG, PEG10000-ICG and PEG20000-ICG in the near infrared I region (650nm-900nm range). The fluorescence intensity.
  • the test results are shown in Figure 4.
  • the weighted continuous curve represents PEG20000-ICG
  • the gray continuous line represents ICG
  • the dark heavy dotted line represents PEG500-ICG
  • the thin dotted line represents PEG2000-ICG
  • the one-dot chain line represents PEG5000-ICG
  • the two-dot chain line represents PEG10000-ICG.
  • the modified products PEG500-ICG, PEG2000-ICG, PEG5000-ICG, PEG10000-ICG, and PEG20000-ICG have good fluorescent properties comparable to unmodified ICG. It can be seen that the modification of ICG by modified PEG did not affect the fluorescent properties of the original ICG.
  • NIRQuest512 and CVH100 / M set the excitation light at 808nm, and test ICG, PEG500-ICG, PEG2000-ICG, PEG5000-ICG, PEG10000-ICG, and PEG20000-ICG in the near infrared II region (in the range of 900nm-1200nm) Relative fluorescence intensity.
  • the test results are shown in Figure 5.
  • the weighted continuous curve represents PEG20000-ICG
  • the gray continuous line represents ICG
  • the dark heavy dotted line represents PEG500-ICG
  • the thin dotted line represents PEG2000-ICG
  • the one-dot chain line represents PEG5000-ICG
  • the two-dot chain line represents PEG10000-ICG.
  • the modified products PEG500-ICG, PEG2000-ICG, PEG5000-ICG, PEG10000-ICG, and PEG20000-ICG have good fluorescent properties comparable to unmodified ICG.
  • ICG modified by mPEG-NH 2 has good fluorescence properties in the near-infrared I and II regions, and is consistent with the absorption and emission spectral properties of ICG. Therefore, PEG-ICGs have the application value of near-infrared I and II region imaging equivalent to ICG.
  • the components of the PBS buffer solution are Na 2 HPO 4 and NaH 2 PO 4 .
  • a spectrophotometer 200 was used, and the excitation light was set at 808 nm.
  • the test results are shown in Figure 6.
  • the results show that except for the decrease in the fluorescence intensity of PEG500-ICG, which is slightly faster within 1 to 12 hours, the fluorescence intensity of other PEG2000-ICG, PEG5000-ICG, PEG10000-ICG, and PEG20000-ICG are stable over time .
  • This shows that the modified products PEG500-ICG, PEG2000-ICG, PEG5000-ICG, PEG10000-ICG, and PEG20000-ICG have good stability in a near neutral environment.
  • the modified products PEG500-ICG, PEG2000-ICG, PEG5000-ICG, PEG10000-ICG, and PEG20000-ICG prepared in Preparation Example 1-5 were respectively dissolved in FBS (fetal bovine serum, SR01C-500 purchased from SAILY BIO) to obtain 5 test samples (20 ⁇ g / mL).
  • FBS fetal bovine serum, SR01C-500 purchased from SAILY BIO
  • mice Eighteen healthy mice (about 20 g) were divided into 6 groups of 3 mice each.
  • the 6 groups of mice were respectively injected with ICG and PEG500 in the tail vein according to the amount of ICG in the composition of 1 mg ⁇ kg -1 (the injection amount was 0.1-1 mg ⁇ kg -1 , and the concentration of ICG in the injection solution was 20 ⁇ g / mL).
  • the blood clearance rates of ICG and the above five mPEG-ICG were compared by near-infrared region II fluorescence imaging and vascular fluorescence imaging.
  • the specific operation method and calculation are as follows:
  • Imaging and photographing conditions Excited by a 808nm laser, and a 900nm long-pass filter FELH0900 separates the excitation light from the emitted light, and is photographed by a SWIR near-infrared camera.
  • mice were anesthetized with the animal anesthesia system VIP3000, and five kinds of PEG-ICGs (PEG500-ICG, PEG2000-ICG, PEG5000-ICG, PEG10000-ICG, and PEG20000-ICG) and ICG were injected into the tail vein at the same time point for 3 min.
  • PEG-ICGs PEG500-ICG, PEG2000-ICG, PEG5000-ICG, PEG10000-ICG, and PEG20000-ICG
  • ICG ICG were injected into the tail vein at the same time point for 3 min.
  • 5min, 6min, 7min, 8min, and 12min near-infrared region II fluorescence imaging was performed on the femoral vein of the mouse, and pictures were taken, and see the attached photo group shown in FIG. 8.
  • the calculation of ICG or PEG-ICGs half-life in plasma is to compare the fluorescence intensity of the blood vessel site in the image photograph with the fluorescence intensity of the skin of the leg to find the ratio to reflect the blood metabolism time of PEG-ICGs and ICG.
  • the blood vessels in the medial leg of the leg were clearly visible 3 minutes after the injection, and the fluorescence signal of the blood vessel part disappeared around 8 minutes.
  • the IC ( T (1/2) 0.06421h ) was calculated.
  • EPR effect that is, the high permeability and retention effect of solid tumors. Specifically, it refers to the property that certain sizes of molecules or particles tend to aggregate in tumor tissue compared to normal tissue. The main reason is that the microvascular endothelial space in normal tissues is dense and structurally complete, and macromolecules and lipid particles cannot easily penetrate the vascular wall, while solid tumor tissue is rich in blood vessels, wide vascular wall space, poor structural integrity, and lack of lymphatic reflux, resulting in Macromolecular substances and lipid particles have selective high permeability and retention. This phenomenon is called the high permeability and retention effect of solid tumor tissues, which is referred to as the EPR effect. An important indicator for measuring the suitability of drugs or diagnostic materials for tumor detection is the EPR effect.
  • the EPR effect of the PEG-ICGs prepared by the present invention was tested. The test method is as follows:
  • skov-3 tumor cells purchased from EK-Bioscience
  • 2 mice weighing 300 g / mouse.
  • Each tumor has a population of 2,000,000 cells and is injected subcutaneously. After 3-4 weeks, the tumor size reaches 5-10 mm.
  • Two tumor-bearing mice were anesthetized by the animal anesthesia system VIP3000, based on the amount of ICG (due to the different molecular weights of ICG and PEG5000-ICG, here 1mg ⁇ kg -1 was calculated based on the amount of ICG contained; injection At that time, ICG and PEG5000-ICG were dissolved in PBS at pH 7.4), and ICG and PEG5000-ICG were injected into the tail vein at 1 mg ⁇ kg -1 each.
  • ICG-injected mice were imaged by near-infrared region II at 3min, 4h, and 6h, and PEG5000-ICG-injected mice were imaged by near-infrared region II at 3min, 12h, and 48h.
  • Imaging and photographing conditions Excited by a 808nm laser, and a 900nm long-pass filter FELH0900 separates the excitation light from the emitted light, and is photographed by a SWIR near-infrared camera. The photos taken are shown in Figure 9.
  • mice injected with PEG5000-ICG the blood vessels at the tumor were very clear after 3 minutes, the fluorescence signal at the tumor was obvious at 12 hours, and there was a strong EPR effect, which was clearer than that of mice injected with IEG at 3 minutes.
  • Imaging of PEG5000-ICG-injected mice after 48h was close to imaging of IEG-injected mice at 3min. It can be seen that the PEG-ICGs of the present invention have a strong EPR effect and can be applied to the detection of tumors in animals.
  • modified PEGs have properties similar to mPEG-NH 2 , such as mPEG, mPEG-SS, mPEG-SC, mPEG2-NHS, mPEG-SPA, mPEG-ALD, mPEG-MAL , HO-PEG-COOH, mPEG-b-PS, mPEG-b-PI, mPEG-b-PAN, mPEG-b-PCL, mPEG-b-PMMA, ⁇ -hydroxyl-PEG- ⁇ -amide, HO-PEG -OH, HO-PEG-NH 2 , mPEG-carboxyl, mPEG-cyan, etc.
  • these modified PEGs can also be used to modify or modify ICG.
  • the process / conditions of the modification reaction may be all different due to the structure of the modified PEG molecule or the end groups, but it will be understandable to those skilled in the art based on the facts and product properties tested and confirmed by the present invention.
  • Some ICGs modified by other modified PEGs should also have half-life and blood metabolic properties similar to those of ICGs modified by mPEG-NH 2 , can stay in the blood for a longer time, and also show the same as unmodified ICGs.
  • Equivalent / even better fluorescence properties and spectral absorption capabilities can be used in angiography, intraoperative navigation, tumor detection, etc., to optimize the application performance of ICG.

Landscapes

  • Health & Medical Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Veterinary Medicine (AREA)
  • Engineering & Computer Science (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Animal Behavior & Ethology (AREA)
  • General Health & Medical Sciences (AREA)
  • Public Health (AREA)
  • Biomedical Technology (AREA)
  • Epidemiology (AREA)
  • General Chemical & Material Sciences (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Medicinal Chemistry (AREA)
  • Polymers & Plastics (AREA)
  • Organic Chemistry (AREA)
  • Medicines Containing Antibodies Or Antigens For Use As Internal Diagnostic Agents (AREA)
  • Medicinal Preparation (AREA)
  • Investigating, Analyzing Materials By Fluorescence Or Luminescence (AREA)

Abstract

La présente invention concerne un vert d'indocyanine modifié, son procédé de préparation et son utilisation. L'un de deux groupes sulfoniques de la structure moléculaire du vert d'indocyanine est substitué par un groupe PEG modifié, obtenant ainsi un nouveau type de vert d'indocyanine (ICG) modifié par du PEG. En modifiant l'ICG, la demi-vie plasmatique de l'ICG peut être prolongée, de sorte que son temps de rétention dans le sang est notablement prolongé, et ainsi la performance d'utilisations dans de multiples aspects, tels que l'angiographie, la navigation intraopératoire, et la détection et le traitement de tumeurs, est optimisée. L'ICG modifié présente de bonnes propriété fluorescente et capacité d'absorption spectrale similaires à l'ICG non modifié d'origine ; en outre, l'ICG modifié présente une très bonne stabilité, et possède par conséquent de bonnes perspectives d'applications dans le domaine de l'imagerie vasculaire.
PCT/CN2018/106522 2018-08-22 2018-09-19 Vert d'indocyanine modifié et son procédé de préparation et son utilisation Ceased WO2020037752A1 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
CN201810959509.7A CN109054013A (zh) 2018-08-22 2018-08-22 一种经修饰的吲哚菁绿及其制备方法
CN201810959509.7 2018-08-22

Publications (1)

Publication Number Publication Date
WO2020037752A1 true WO2020037752A1 (fr) 2020-02-27

Family

ID=64687962

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/CN2018/106522 Ceased WO2020037752A1 (fr) 2018-08-22 2018-09-19 Vert d'indocyanine modifié et son procédé de préparation et son utilisation

Country Status (2)

Country Link
CN (1) CN109054013A (fr)
WO (1) WO2020037752A1 (fr)

Families Citing this family (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN112898578A (zh) * 2020-02-26 2021-06-04 南昌大学第一附属医院 一种吲哚菁绿衍生物制备方法及其在检测氧化型低密度脂蛋白中的应用
CN112778515B (zh) * 2021-01-29 2023-03-14 佛山市华联有机硅有限公司 一种聚醚型磺酸及其无溶剂制备方法与应用

Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2014185312A (ja) * 2012-07-20 2014-10-02 Canon Inc 化合物、および、前記化合物を有する光音響イメージング用造影剤
CN104548139A (zh) * 2014-12-18 2015-04-29 深圳先进技术研究院 一种光声成像用纳米复合造影剂及其制备方法
CN106729769A (zh) * 2017-02-14 2017-05-31 天津医科大学 Tem1特异性荧光探针及其应用
CN108130069A (zh) * 2017-12-27 2018-06-08 深圳大学 稀土上转换纳米诊疗剂及其制备方法

Family Cites Families (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2011506673A (ja) * 2007-12-14 2011-03-03 バイオティウム， インコーポレイテッド 蛍光性化合物
BRPI1009346A2 (pt) * 2009-03-19 2017-05-30 General Eletric Company metodo de formacao de imagem optica in vivo das margens tumorais de um tumor, agente de contraste, composicao farmaceutica, e, kit
CN104470546B (zh) * 2012-07-20 2018-02-27 佳能株式会社 化合物和含有所述化合物的光声成像造影剂
US10245329B2 (en) * 2014-09-08 2019-04-02 Canon Kabushiki Kaisha Composition having dye and conjugate of polyethyleneglycol and additive and contrast agent for photoacoustic imaging having the same
CN105194670A (zh) * 2015-10-28 2015-12-30 温州医科大学 一种阳离子聚合物负载紫杉醇/吲哚菁绿共传输胶束及其制备方法
CN105920625B (zh) * 2016-04-19 2019-02-05 中山大学 一种集超声造影与热疗于一体的诊疗制剂的制备方法

Patent Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2014185312A (ja) * 2012-07-20 2014-10-02 Canon Inc 化合物、および、前記化合物を有する光音響イメージング用造影剤
CN104548139A (zh) * 2014-12-18 2015-04-29 深圳先进技术研究院 一种光声成像用纳米复合造影剂及其制备方法
CN106729769A (zh) * 2017-02-14 2017-05-31 天津医科大学 Tem1特异性荧光探针及其应用
CN108130069A (zh) * 2017-12-27 2018-06-08 深圳大学 稀土上转换纳米诊疗剂及其制备方法

Also Published As

Publication number Publication date
CN109054013A (zh) 2018-12-21

Similar Documents

Publication Publication Date Title
Yang et al. cRGD-functionalized, DOX-conjugated, and 64Cu-labeled superparamagnetic iron oxide nanoparticles for targeted anticancer drug delivery and PET/MR imaging
Yuan et al. Peptide-based semiconducting polymer nanoparticles for osteosarcoma-targeted NIR-II fluorescence/NIR-I photoacoustic dual-model imaging and photothermal/photodynamic therapies
Marshall et al. Single-dose intravenous toxicity study of IRDye 800CW in Sprague-Dawley rats
Huang et al. A porphyrin-PEG polymer with rapid renal clearance
Wang et al. A theranostic nanoplatform: magneto-gold@ fluorescence polymer nanoparticles for tumor targeting T 1 & T 2-MRI/CT/NIR fluorescence imaging and induction of genuine autophagy mediated chemotherapy
US8753608B2 (en) Complex and contrast agent for photoimaging using the same
Liu et al. Croconaine-based nanoparticles enable efficient optoacoustic imaging of murine brain tumors
CN111298140B (zh) 还原响应的t1/t2切换型mri造影剂、其制备方法及应用
Hu et al. Tailoring albumin-based theranostic PROTACs nanoparticles for enhanced NIR-II bioimaging and synergistic cancer chemo-phototherapy
Huang et al. Sericin-based gadolinium nanoparticles as synergistically enhancing contrast agents for pH-responsive and tumor targeting magnetic resonance imaging
US20230158176A1 (en) Near-infrared fluorescent small-molecule probe, synthesis method and application thereof
US20170202980A1 (en) Method for producing optical imaging contrast agent and optical imaging contrast agent
WO2015039555A1 (fr) Agent de contraste à base de matériau d'oxyde de graphène et procédé de préparation associé
JPH06181890A (ja) Ｍｒｉ造影剤
Han et al. Facile synthesis of zwitterionic polyglycerol dendrimers with a β-cyclodextrin core as MRI contrast agent carriers
EP2604289B1 (fr) Forme radiale de composé polymère contenant de l'iode, procédé de préparation de celui-ci, et composition d'agent de contraste tdm contenant celle-ci
Magalotti et al. Evaluation of inflammatory response to acute ischemia using near-infrared fluorescent reactive oxygen sensors
JP2024508128A (ja) 注射用の新規アビラテロン誘導体の調製方法及び用途
WO2020037752A1 (fr) Vert d'indocyanine modifié et son procédé de préparation et son utilisation
Wu et al. Multi-modal imaging for uptake of peptide ligand specific for CD44 by hepatocellular carcinoma
Botz et al. Hydrophobic cyanine dye-doped micelles for optical in vivo imaging of plasma leakage and vascular disruption
KR102014159B1 (ko) 요오드를 포함하는 생체적합성 단분자 담도 컴퓨터 단층촬영용 조영제 및 이의 제조방법
Li et al. Melanin theranostic nanoplatform as an efficient drug delivery system for imaging-guided renal fibrosis therapy
Desu et al. Non-invasive detection of lung inflammation by near-infrared fluorescence imaging using bimodal liposomes
KR102041246B1 (ko) 양쪽이온성 알긴산 유도체 및 이를 포함하는 조영제 조성물

Legal Events

Date Code Title Description
121 Ep: the epo has been informed by wipo that ep was designated in this application

Ref document number: 18930953

Country of ref document: EP

Kind code of ref document: A1

NENP Non-entry into the national phase

Ref country code: DE

122 Ep: pct application non-entry in european phase

Ref document number: 18930953

Country of ref document: EP

Kind code of ref document: A1