Food ingredients; pharmaceutical excipient
Trehalose targets the autophagy pathway. It induces autophagy through an mTOR-independent mechanism. Trehalose activates TFEB (Transcription Factor EB), a master controller of lysosomal biogenesis and autophagy, by inhibiting AKT which is a negative regulator of TFEB. This promotes autophagy by increasing the production of membrane proteins, lysosomal degradation enzymes, and other autophagy-related components. Trehalose also functions as a chemical chaperone, stabilizing misfolded proteins.

In vitro, trehalose acts as a potent chemical chaperone, stabilizing misfolded proteins and preventing aggregation . It induces autophagy in various mammalian cell lines and upregulates TFEB (Transcription Factor EB), leading to enhanced lysosomal biogenesis and autophagic flux . In cellular models, trehalose has been shown to clear mutant proteins (such as huntingtin and α-synuclein) and protect retinal pigment epithelial cells from oxidative damage .

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Typical cocoa substances like D-trehalose can create a special protective film on the surface of cells to protect them from extremes in temperature, high or low humidity, high osmotic pressure, dryness, and water loss. This protects the biological features and life processes of living things. 1. Stabilize the structure of proteins and membranes 2. Avoid drying out 3. First-aid ingredients, pharmaceutical excipients 4. The adsorption impact of cytokine collagen on tube wall proteins is evident, making it challenging for the recombinant protein modification to separate on the tube wall. Consequently, the concentration in the solution is low, and the activity eventually declines. Recombinant proteins that cannot be used for serum-free culture, in vivo experiments, or protein labeling can be protected during reconstitution by adding egg carriers in conjunction with the protein anchoring points on the pre-sealed plastic tube wall to prevent the recombinant protein from appearing on the tube wall. contains human or animal proteins such HSA, FBS, or BSA; in the absence of these, trehalose can be utilized as a carrier protein to keep recombinant proteins for a long time. Protein instability must be prevented at a specific pH and salt concentration. It is not advised to use water to dissolve the carrier. For this solution, a buffer with a pH close to neutral and a specific amount of buffering power, like PBS or culture medium (RPMI1640, DMEM), is typically advised.
In vitro, trehalose acts as a potent chemical chaperone, stabilizing misfolded proteins and preventing aggregation. It induces autophagy in various mammalian cell lines and upregulates TFEB, leading to enhanced lysosomal biogenesis and autophagic flux. Trehalose treatment increases autophagic flux without affecting lysosomal acidification or cell viability. Trehalose induces autophagy in LNCaP cells, detected by quantification of LC3-II/LC3-I ratio. It is a novel mTOR-independent autophagy enhancer that accelerates the clearance of mutant huntingtin and alpha-synuclein.

In animal models, trehalose has been shown to cross the blood-brain barrier and exert therapeutic effects in various diseases . In a rat model of type 2 diabetes, intraperitoneal injection of trehalose increased serum total antioxidant capacity and superoxide dismutase activity . Oral trehalose reduced neuropathology in transgenic mouse models of Huntington's and Alzheimer's diseases . Furthermore, in a mouse model of Mucopolysaccharidosis type II, oral trehalose reduced GAG accumulation, inflammation, and apoptosis in the brain, while improving cognitive function .
In vivo, trehalose protects cells against numerous environmental stresses. It has been shown to remodel human microbiota in gut models, reducing Clostridium difficile infection and promoting the growth of competitive bacteria. Trehalose is neuroprotective in in vitro models of glaucoma. It is used as a food ingredient and active molecule excipient.

In vitro enzyme or receptor binding (non-cell) assays for trehalose are not typically performed, as it is not a direct enzyme inhibitor or receptor ligand. Its autophagy-inducing activity is assessed in cell-based assays. The compound's ability to stabilize proteins can be studied using biochemical assays with purified proteins, where protein aggregation is measured in the presence of trehalose.

In vitro cell-based assays for trehalose involve treating cells with trehalose at concentrations ranging from 20-100 mM. Autophagy is detected by quantification of LC3-II/LC3-I ratio via Western blotting. Autophagic flux is assessed using bafilomycin A1 (Baf) or chloroquine (CQ) to block lysosomal degradation. The presence of autophagic vesicles is analyzed by transmission electron microscopy (TEM). Cell viability is assessed using MTT assays.

In vivo animal experiments for trehalose are conducted using various animal models. Trehalose has been studied for its effects on Clostridium difficile infection in gut models. Neuroprotective effects have been evaluated in models of glaucoma. Trehalose is administered orally or intravenously in toxicity studies. The compound's effects on autophagy and protein clearance are evaluated in tissue samples.

Exogenous trehalose is hydrolyzed into two glucose molecules by the membrane-bound enzyme trehalase in the intestine before absorption . While endogenous human trehalase activity is generally low, low doses are completely digested, whereas high doses may result in some trehalose reaching the colon for microbial fermentation . Intravenously administered trehalose is primarily excreted unchanged by the kidneys .
Pharmacokinetic (PK) properties of trehalose indicate that it has a molecular weight of 342.30 and a molecular formula of C12H22O11. It is a naturally occurring disaccharide. Trehalose is hydrolyzed by trehalase in the small intestine to glucose. It is well-tolerated and has low systemic toxicity. The compound is widely used as a food ingredient and excipient. Storage at room temperature is recommended for the anhydrous form.

Trehalose is considered extremely safe. Extensive evaluations, including multiple animal toxicity studies, have found no consistent treatment-related adverse effects, even at dietary doses as high as 10% . Human studies indicate that it is well-tolerated at daily doses up to 50 grams, with only mild gastrointestinal symptoms (such as bloating or diarrhea) possible at high doses due to its incomplete absorption . It is approved as Generally Recognized as Safe (GRAS) by the FDA and is widely used in the food and pharmaceutical industries.
Toxicology (toxicology) data for trehalose indicate that it is safe for use as an ingredient in consumer products. Acute toxicity studies in mice, rats, and dogs showed no mortalities due to trehalose treatments. A single oral dose of 5 g/kg or intravenous dose of 1 g/kg showed no systemic toxicity in rats and mice. No consistent treatment-related, dose-dependent adverse effects were observed in eight safety studies at doses up to 10% of the diets. Trehalose did not induce maternal or developmental toxicity at concentrations up to 10% in the diet.

[1]. Amorphous trehalose dihydrate by cryogenic milling. Carbohydr Res. 2011 Jun 1;346(8):1061-4.

α,α-trehalose is a type of trehalose in which both terminal carbons of the glucose residues are in the α-configuration. It is a metabolite found in humans, Saccharomyces cerevisiae, Escherichia coli, and mice, and possesses anti-aging properties. Cabaletta has been used in clinical trials for the treatment of oculopharyngeal muscular dystrophy. Trehalose is a metabolite found or produced in Escherichia coli (K12 strain, MG1655 strain). Trehalose has also been reported in Microchloropsis, Myrothamnus flabellifolia, and other organisms with relevant data. Trehalose is a metabolite found or produced in Saccharomyces cerevisiae.
Other information: Trehalose is a naturally occurring non-reducing disaccharide. It is widely used as a food ingredient and active molecule excipient. Trehalose is an inducer of autophagy and protects cells against environmental stresses. It is also known as D-(+)-Trehalose. Its CAS number is 99-20-7.

C12H22O11

342.297

342.116

C, 42.11; H, 6.48; O, 51.41

99-20-7

7427

White to off-white solid powder

1.8±0.1 g/cm3

675.4±55.0 °C at 760 mmHg

203 °C

362.3±31.5 °C

0.0±4.7 mmHg at 25°C

1.652

-3.3

8

11

4

23

348

10

C([C@@H]1[C@H]([C@@H]([C@H]([C@H](O1)O[C@@H]2[C@@H]([C@H]([C@@H]([C@H](O2)CO)O)O)O)O)O)O)O

HDTRYLNUVZCQOY-LIZSDCNHSA-N

InChI=1S/C12H22O11/c13-1-3-5(15)7(17)9(19)11(21-3)23-12-10(20)8(18)6(16)4(2-14)22-12/h3-20H,1-2H2/t3-,4-,5-,6-,7+,8+,9-,10-,11-,12-/m1/s1

(2R,3S,4S,5R,6R)-2-(hydroxymethyl)-6-[(2R,3R,4S,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)oxan-2-yl]oxyoxane-3,4,5-triol

NSC-2093; trehalose; 99-20-7; D-(+)-Trehalose; Mycose; alpha,alpha-trehalose; NSC 2093; Trehalose

2934.99.9001

Powder      -20°C    3 years

                     4°C     2 years

In solvent   -80°C    6 months

                  -20°C    1 month

Note: Please store this product in a sealed and protected environment (e.g. under nitrogen), avoid exposure to moisture.

Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)

H2O : ~250 mg/mL (~730.35 mM)

Solubility in Formulation 1: 100 mg/mL (292.14 mM) in PBS (add these co-solvents sequentially from left to right, and one by one), clear solution; with sonication.

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 (Please use freshly prepared in vivo formulations for optimal results.)