Table 4 Physicochemically targeted drug delivery system for norcantharidin

NCTD-loaded thermosensitive in-situ gel

28% poloxamer 407, 1.6% poloxamer 188, 0.1% hydroxypropyl methyl cellulose (HPMC), NCTD

Thermosensitive: gelation temperature: 34 ℃, dissolution time: 210 min; sustained-release property in vitro

-

[145], 2017

NCTD-loaded thermosensitive in-situ gel

62.5 mg NCTD, 7 g poloxamer 407, 0.4 g poloxamer 188, 0.025 g HPMC

Thermosensitive: gelation temperature and time: (33.9–34.1) ℃, (101–103) s; viscosity: > 100,000 mPa·s (37 ℃); sustained-release property in vitro

H22 tumor inhibition rate: NCTD (ip., once daily, 2 mg/kg) < NCTD-loaded thermosensitive in-situ gel (intratumoral injection, once every 3 days, 3.3 mg/kg) < NCTD-loaded thermosensitive in-situ gel (intratumoral injection, once every 3 days, 6.6 mg/kg) < NCTD-loaded thermosensitive in-situ gel (intratumoral injection, once every 3 days, 9.9 mg/kg)

[146], 2017

NCTD-loaded thermosensitive in-situ gel

62.5 mg NCTD, 7 g poloxamer 407, 0.4 g poloxamer 188, 0.025 g HPMC

Thermosensitive: gelation temperature: 34 ℃, dissolution time: 3.5 h

H22 tumor inhibition rate: NCTD (ip., once daily, 2 mg/kg) < NCTD-loaded thermosensitive in-situ gel (intratumoral injection, once every 3 days, 3.3 mg/kg) < NCTD-loaded thermosensitive in-situ gel (intratumoral injection, once every 3 days, 6.6 mg/kg) < NCTD-loaded thermosensitive in-situ gel (intratumoral injection, once every 3 days, 9.9 mg/kg); survival time ↑; inhibit VEGF and CD44 expression ↑

[147], 2019

NCTD-loaded metal-organic framework (IRMOF-3) coated with a poloxamer thermosensitive gel (NCTD-IRMOF-3-Gel)

15 mg NCTD-IRMOF-3 (NCTD, IRMOF-3), 3 mL freeze-dried protective agent (4% mannitol and 2% poloxamer)

Thermosensitive; NCTD release (5 h): sustained-release effect: NCTD (90% NCTD) > NCTD-IRMOF-3 (50% NCTD) > NCTD-IRMOF-3-Gel (30% NCTD)

Hepa1-6 cell: cell inhibition rate: NCTD-IRMOF-3-Gel > NCTD, NCTD-IRMOF-3; block cell cycle in the S and G2/M phases; cell apoptosis (48 h, 50 µg/mL): NCTD-IRMOF-3-Gel > NCTD-IRMOF-3 > NCTD

[148], 2021

Thermosensitive

hydrogel co-loaded with NCTD nanoparticles and doxorubicin (NCTD-NPs/Dox Gel)

85 mg NCTD-NPs (PCEC copolymers, NCTD), 2 mg Pluronic F127 (PF127), 30 mg Dox

Thermosensitive: gelation temperature: 29.4 ℃; NCTD release: sustained-release effect: NCTD (48 h, 94.2% NCTD) > NCTD-NPs (168 h, 84.5% NCTD) > NCTD-NPs/Dox Gel (168 h, 47.8% NCTD)

HepG2 cells: IC₅₀: NCTD-NPs/Dox Gel < free NCTD/Dox; H22 tumor inhibition rate (intratumoral injection, 0.1 mL, once a week for two times): free NCTD/Dox (2.5 mg/kg free NCTD and 10 mg/kg free Dox) < NCTD-NPs/Dox Gel (equivalent to

2.5 mg/kg free NCTD and 10 mg/kg free Dox); median

survival time: free NCTD/Dox group (53 days) < NCTD-NPs/Dox Gel (67 days); no visible tissue damage, inflammation, or lesions; inhibit tumor proliferation and angiogenesis: Ki-67-positive cells: NCTD-NPs/Dox Gel (22.46 ± 2.51) % < free NCTD/Dox (38.77 ± 4.85) %, CD31-positive microvessel density (MVD): NCTD-NPs/Dox Gel (7.67 ± 0.94) < free NCTD/Dox (15.34 ± 2.05)

[149], 2021

Lactosyl-NCTD (Lac-NCTD) phospholipid complex (LPC) loaded liposome (pH-LPC-lips)

40 mg soybean phosphatidylcholine, 10 mg cholesterol, 4 mg Lac-NCTD LPC, 2 mL 0.02% CMCT solution

pH sensitivity: cumulative release of NCTD: pH = 5.2 > pH = 6.8 > pH = 7.4

-

[152], 2011

pH-LPC-lips

40 mg soybean phosphatidylcholine, 10 mg cholesterol, 4 mg Lac-NCTD LPC, 2 mL 0.02% CMCT solution

pH sensitivity

HepG2 cells: IC₅₀: pH-LPC-lips (0.094 µmol/mL) < Lac-lips (0.140 µmol/mL) < Lac-NCTD (0.351 µmol/mL); cellular uptake: pH-LPC-lips > Lac-lips > Lac-NCTD; H22 tumor inhibition rate (ip., once daily, 10 days): NCTD (2.0 mg/kg) = 31.2% < Lac-NCTD (6.6 mg/kg) = 38.3% < Lac-lips (6.6, 13.2 mg/kg) = 56.6%, 76.9% < pH-LPC-lips (6.6, 13.2 mg/kg) = 63.6%, 85.3%; the accumulation of NCTD in liver and tumor tissues ↑

[153], 2014

Cross-linked polymer nano-cooperative prodrugs (PPD-NPs)

PPD [cisplatin/NCTD dual drug small molecule (Pt (IV)-1), EDC, NHS, DMF, polyorthoester main chain (POEAd-NH₂)], DMSO, phosphate buffer (pH = 7.4, 50 mM)

pH sensitivity: Pt release (24 h): pH = 5.0 + 10 mM GSH (80%) > pH = 6.8 (30%) > pH = 7.4 (15%); H22 and HepG2 cellular uptake: pH = 6.8 > pH = 7.4

H22 and HepG2 cells: IC₅₀: PPD-NPs (pH = 6.8) < PPD-NPs (pH = 7.4) < cisplatin + NCTD < Pt (IV)-1; induce cell apoptosis; concentration of Pt in mouse blood and tumors: PPD-NPs > Pt (IV)-1 > cisplatin + NCTD; H22 tumor inhibition rate (iv., 6 mg/kg): PPD-NPs > Pt (IV)-1 > cisplatin + NCTD; Pt-DNA ↑, PPA2 activity ↓; inhibit HepG2 tumor growth (PPD-NPs); heart and kidney toxicity ↓

[156], 2021

NCTD-loaded TPP-PEG-PCL nanomicelles (NCTD@TPP-

PEG-PCL)

10 mg NCTD, 100 mg TPP-PEG-PCL, PBS

Attraction of positive and negative charges: TPP cations (positive charge), cell membranes and mitochondrial membranes (negative charge)

NCTD@TPP-PEG-PCL/NCTD@PEG-PCL/NCTD: t_1/2, AUC, MRT ↑; SMMC-7721 cells: cellular uptake ↑, mitochondrial targeting ability ↑, lysosome escape ↑; inhibit cell growth and induce cell apoptosis: NCTD@TPP-PEG-PCL > NCTD@PEG-PCL > NCTD; mitochondrial membrane potential ↓, ROS ↑, Bcl-2 ↓, Bax ↑

[159], 2020

Charge-

reversal polymer nano-modulator (SP_DMCN)

The amphiphilic semiconducting polymer with acid-labile DMC-incorporating conjugates on the surface (SP_DMC)

pH sensitivity and charge conversion: zeta potential: -17 mV (pH 7.4), + 7 mV (pH 6.5), + 12 mV (pH 5.0); ¹O₂ generation: pH 6.7 > pH 7.4; tumor penetration ↑

CT26 cells: SP_DMCN (IC₅₀ = 22 µg/mL) < SP_SAN (IC₅₀ = 30.8 µg/mL); PP2A ↓, PI3K/AKT ↑, mTORC1 ↑, FOXP3 ↓, Treg ↓;

CT26 primary and distant tumor inhibition rate (iv., 2 mg [SPN]/kg, laser condition: 808 nm, 0.3 W/cm², 8 min): SP_DMCN > SP_SAN; caspase-3 ↑; favorable in vivo biosafety and biocompatibility; CD8⁺/CD4⁺ ratio ↑; granzyme B ↑; HMGB1 (ICD) ↑; CD11c⁺CD80⁺CD86⁺ DCs (DC maturation) ↑; IFN-γ, IL-6, TNF-α ↑; CD4⁺CD25⁺FOXP3⁺ Tregs cells among CD3⁺ T cells ↓; CD8⁺/Treg ratio ↑; CTL ↑

[160], 2021

Light-activatable dual prodrug polymer nanoparticle (DPP NP)

DP monomer (100 mg, 1.0 eq), EDC (58.8 mg, 2.3 eq), NHS (35.5 mg, 2.3 eq), DMF (8 mL), EDA (7.9 mg, 0.98 eq), mPEG_2 K-NH₂ (20 mg, 0.08 eq)

Light responsiveness: drug release could be precisely controlled by visible light (upon periodic irradiation and upon preirradiation)

HeLa cells: cellular uptake ↑, p-AKT protein and Pt-DNA adducts (light irradiation) ↑, IC₅₀ (light irradiation): DPP NP (53.5 µM) < DP (64.1 µM) < NCTD (108 µM) < Pt(IV) (170 µM); KM mice bearing U14 tumor: Pt DMCT imaging of DPP NP; U14 tumor inhibition rate (iv., 3 mg Pt/kg on days 0, 3 and 6, light irradiation: 430 nm, 20 mW/cm², 30 min): 75% of mice were fully cured, apoptosis ↑, survival rate ↑

[162], 2019

NCTD-loaded PLGA-alginate microsphere (NPAM)

2 mL ethyl acetate containing 0.5 g PLGA and 0.3 g NCTD, 50 g aqueous solution containing sodium alginate, 75 g isooctane containing 2.54 g Span 85, 5 g aqueous solution containing 1.36 g Tween 85, 20 g 15% (w/w) calcium chloride solution

Chemoembolization; average size: (46.9 ± 5.4) µm (PLGA: alginate = 1: 3, w: w); drug released by burst effect ↓; disintegration time: about 4 days

SMMC-7721 cells: NPAM: IC₅₀ (24 h, 48 h, 72 h) = 110.2, 70.6, 35.5 µg/mL; the growth rate of the tumor after treatment: 1.5 mL/kg 0.03% (w/v) NCTD solution (12.4) > 10 mg/kg blank PLGA-alginate microspheres (BPAM) (10.1) > 10 mg/kg NPAM (1.1); survival time and survival rate: 1.5 mL/kg 0.03% (w/v) NCTD solution (15.8 ± 2.0 days, 15.8%) < 10 mg/kg BPAM (16.5 ± 3.0 days, 20.7%) < 10 mg/kg NPAM (31.0 ± 3.9 days, 126.8%)

[165], 2006

NCTD-chitosan microsphere (NCTD-CS-MS)

NCTD, 5% dilute acetic acid, CS, liquid paraffin containing Span-80, 25% glutaraldehyde-saturated toluene solution

Chemoembolization; average size: (143.54 ± 4.24) µm, within 60–200 μm account for 87%; sustained-release property (cumulative release rate in 7 days reaches about 60%)

Establishment of a rabbit VX-2 liver cancer model by ultrasound-guided puncture (NCTD: 1.0 mg/kg, NCTD-CS-MS: 10.09 mg/kg): survival time and life prolonging rate: NCTD (16.23 ± 0.45 days, 20.69 ± 3.35%) < NCTD-CS-MS (25.73 ± 0.60 days, 91.33 ± 4.48%); tumor growth rate: NCTD-CS-MS (9.72%) < NCTD (12.9%)

[166], 2010

NCTD sustained-release microsphere (NCTD-MS)

NCTD, 10 mL 2.0% sodium alginate, 0.15 g nanoscale calcium carbonate, 50 mL liquid paraffin containing 1% Span-80, 1.0 mL glacial acetic acid, 2% CS solution; drug/carrier (sodium alginate + calcium carbonate) = 0.8:1 (w/w)

Chemoembolization; average size: (309.75 ± 2.19) µm; sustained-release property; NCTD release: NCTD-MS (normal saline, pH 7.4 PBS) = 54%, 32% (3 h) and 80% (24 h) < NCTD (normal saline, pH 7.4 PBS) = 100% (3 h)

-

[167], 2011

Lipidic solid dispersion of NCTD-loaded

alginate/CS microsphere (LSD/NTCD-ACM)

NCTD, sodium alginate, 50 mL liquid paraffin containing Span-80, CS, calcium chloride, glutaraldehyde, insect wax

Chemoembolization; skeleton type sustained release effect; NCTD release: LSD/NTCD-ACM (120–200 μm) = (53.05 ± 2.73) % (5 days) < LSD/NTCD-ACM (60–120 μm) = (73.65 ± 0.94) % (5 days) < LSD/NTCD = (82.44 ± 1.36) % (5 days) < NCTD-ACM (120–200 μm) = 80% (24 h) < NCTD = 100% (3 h)

Establishment of a rabbit VX-2 liver cancer model by ultrasound-guided biopsy needle method: survival time and life prolonging rate: NCTD [(26.67 ± 0.58) days, -(0.03 ± 4.55) %] < LSD/NTCD-ACM (60–120 μm) [(39.49 ± 0.51) days, (43.81 ± 4.34) %] < LSD/NTCD-ACM (120–200 μm) [(43.37 ± 0.45) days, (57.94 ± 5.76) %]; tumor growth rate: LSD/NTCD-ACM (60–120 μm) = (7.76 ± 0.41) % < LSD/NTCD-ACM (120–200 μm) = (9.56 ± 0.37) % < NCTD = (13.37 ± 1.63) %

[168, 169], 2011, 2013

NCTD-loaded silk fibroin/CS microsphere (NCTD-SF/CS-MS)

NCTD, silk fibroin, CS, dilute acetic acid, liquid paraffin containing Span-80, glutaraldehyde

Chemoembolization; average size: (184 ± 5) µm; sustained-release property; NCTD release: NCTD-SF/CS-MS < NCTD-CS-MS

Establishment of a rabbit VX2 liver tumor model by ultrasound-guided percutaneous puncture: survival time and life prolonging rate: NCTD [(26.67 ± 0.58) days, -0.03%] < blank-microsphere (B-MS) (100–200 μm) [(35.79 ± 0.26) days, 25.83%) < NCTD-CS-MS (100–200 μm) [(37.51 ± 0.46) days, 31.89%] < NCTD-SF/CS-MS (100–200 μm) [(40.29 ± 0.34) days, 41.66%]; tumor growth rate: NCTD-SF/CS-MS (100–200 μm) < NCTD-CS-MS (100–200 μm) < B-MS (100–200 μm) < NCTD; tumor necrosis rate ↑

[172], 2013

NCTD-N-chitosan/silk fibroin microsphere (NCTD-N-CS/SF-MS)

NCTD, silk fibroin, N-CS, dilute acetic acid, liquid paraffin containing Span-80, glutaraldehyde

Chemoembolization; average size: (117 ± 4.3) µm; sustained-release property; NCTD release: NCTD-N-CS/SF-MS = 30% (40 min), 60% (7 days) < NCTD = 100% (40 min)

Establishment of a rabbit VX2 liver tumor model by ultrasound-guided percutaneous puncture: survival time and life prolonging rate: NCTD (23.25 days, -1.25%) < NCTD + N-CS/SF-MS (31 days, 31.91%) < N-CS/SF-MS (34 days, 44.68%) < NCTD-N-CS/SF-MS (36.25 days, 54.25%); tumor inhibition rate and tumor cell necrosis rate: NCTD-N-CS/SF-MS [85.01%, (56.78 ± 0.84) %] > NCTD + N-CS/SF-MS [62.98%, (52.23 ± 0.64) %) > N-CS/SF-MS [58.16%, (49.63 ± 1.02) %] > NCTD [27.22%, (19.35 ± 0.92) %];

[173, 174] 2013, 2014