Carbonic Anhydrase Is A Key Driver Of Aging; Inhibiting It Is Beneficial

[LeeLemonoil]

No, apigenin comes as a glycoside in nature and those are water-soluble.

 

[Inaut]

Phew!!!!

Thanks @LeeLemonoil :)

 

[SQu]

Yep, Chamomille is not one of the staple phytopharmacons for nothing. Apigenin is a gem and Peat advocates it since years

What do you think the most effective way to use it is? Especially if you don't love the tea?

 

[LeeLemonoil]

Eat it. Tea can become an acquired taste, just try to get used to it

 

[LeeLemonoil]

I mean eat the chamomile buds. I buy organic buds for tea but also eat the buds after brewing them, to get all the Apigenin that didn’t go into solution and the many other beneficial substances in it. It’s eccentric I know, but my wife and kids are used to it. And I‘m a healthy bastard so I and justify any of these eccentricities with „he who cures/is in health is right“.
You can just as well eat raw chamomile, it tastes floral-spicy. An interesting taste, put it into a salad

 

[SQu]

I'll give it a bash! Thanks

 

[Sativa]

so i looked up the solubility of apigenin and it is insoluble in water....does that mean i need to eat the parsley/chamomile for its benefit instead of brewing teas? ill test it out

Yup, water will not pull much of the insoluble components. Maybe some, but not all.
I have access to pure German Chamomile CO2 extract, which provides a full spectrum extract. Very potent, both therapeutically and psychoactively.
I also have some pure Apigenin powder lying around somewhere.

 

[Inaut]

I started eating the parsley/chamomile after I brewed it (since yesterday). One way or another I’ll get it in me ‍

 

[Epistrophy]

In light of hearing what molecules can inhibit carbonic anhydrase (CA), I have found a study that puts light into the lesser talked about CA activators. In the study the authors give this reasoning as to why "This topic, then, received little attention from the scientific community for at least two reasons: (i) the statement by Clark and Perrin (1951) that activators of CA do not exist and (ii) the idea that the reported activation is not a phenomenon per se but an artifact generally due to restoration of CA activity possibly lost in the presence of adventitious metal ions or other impurities (or due to enzyme adsorption at interfaces, or even due to enzyme denaturation followed by renaturation in the presence of activators) (Maren, 1967) Leiner (1940), the researcher whose role in discovering this important class of modulators of CA activity should be completely reevaluated, observed among others that the activation was readily detected when working with highly purified enzyme preparations, and this may explain the large discrepancies between the different early studies describing this phenomenon."

The authors describe several activators and their interactions with the CA enzyme which include: histamine, L and D-phenylalanine, L and D-histidine, adduct of human isozyme (hCA) with L-histidine, and L-Adrenaline. This study shines light on the validation of lowering stress hormones and using gelatin as much as possible to negate the effects of some inflammatory amino acids.

 

[methylenewhite]

I started eating the parsley/chamomile after I brewed it (since yesterday). One way or another I’ll get it in me ‍

Do you still do it? Any results?

 

[Dr. B]

In light of hearing what molecules can inhibit carbonic anhydrase (CA), I have found a study that puts light into the lesser talked about CA activators. In the study the authors give this reasoning as to why "This topic, then, received little attention from the scientific community for at least two reasons: (i) the statement by Clark and Perrin (1951) that activators of CA do not exist and (ii) the idea that the reported activation is not a phenomenon per se but an artifact generally due to restoration of CA activity possibly lost in the presence of adventitious metal ions or other impurities (or due to enzyme adsorption at interfaces, or even due to enzyme denaturation followed by renaturation in the presence of activators) (Maren, 1967) Leiner (1940), the researcher whose role in discovering this important class of modulators of CA activity should be completely reevaluated, observed among others that the activation was readily detected when working with highly purified enzyme preparations, and this may explain the large discrepancies between the different early studies describing this phenomenon."

The authors describe several activators and their interactions with the CA enzyme which include: histamine, L and D-phenylalanine, L and D-histidine, adduct of human isozyme (hCA) with L-histidine, and L-Adrenaline. This study shines light on the validation of lowering stress hormones and using gelatin as much as possible to negate the effects of some inflammatory amino acids.

how do carbonic anhydrase inhibitors affect hydrogen peroxide levels mate

 

[Amazoniac]

I came across the following publication after our ??n's latest class.

- Aspirin: A Suicide Inhibitor of Carbonic Anhydrase II

Spoiler

"In addition to sulfonamides, a variety of other chemical motifs have been identified to inhibit CA, such as carboxylic acids [15]. Nicotinic and ferulic acid have recently been identified as inhibitors of CAII [16]. Unlike the sulfonamide-based drugs, these inhibitors do not directly displace the zinc bound solvent, but instead anchor through the solvent, blocking substrate entry to the active site [16]. Furthermore, 3-nitrobenzoic acid has also been reported as a potent CAI, with further studies showing its potential clinical relevance as a cancer therapeutic [17]. Previous work has also shown that salicylic acid as well as some phenol derivatives are μM inhibitors of mammalian CAs, although the exact mechanism of inhibition for salicylic acid is unknown [18]. These carboxylic acid-based compounds represent a new and largely unstudied class of CAIs."

"A recent study found in a genome-wide search that CAII is the only protein overexpressed in patients with Aspirin resistance and therefore may be the unidentified carboxylesterase [24]. Since Aspirin is a carboxylic acid-based molecule, it was hypothesized that it could potentially bind to CAII. Here, we examine this hypothesis through structure activity relationship studies between Aspirin and CAII, through X-ray crystallography and a spectroscopy-based kinetic assay. We determine that CAII is the previously unidentified carboxylesterase responsible for Aspirin’s short half-life in the blood, and that the product of this reaction, SA, can then inhibit CAII, thus making Aspirin a suicide inhibitor."

"A known and widely studied function of CAs is their ability to act as an esterase [37]. When a small molecule with an ester bond such as Aspirin enters the active site of CAII, its ester bond is cleaved leaving an acetyl group and SA in the case of Aspirin. The zinc bound hydroxide is a strong nucleophile, able to attack the carbonyl of an ester, cleaving the bond."

"Based on the crystallographic data of SA bound to CAII and the modeling with Aspirin, we propose a mechanism for CAII Aspirin ester cleavage and SA inhibition (Figure 4). Firstly, Aspirin binds to the zinc bound solvent within the active site with its acetate bound in a similar fashion of CO2 binding, positioned for nucleophilic attack (Figure 4A). The hydroxyl cleaves the ester bond in the Aspirin molecule leaving the acetate bound to the active site (Figure 4B). The acetate of the reaction is displaced by a water molecule that binds the zinc, while the SA remains in the active site (Figure 4C,D). Finally, the SA reorients within the active site and anchors through the zinc bound solvent, inhibiting any further reaction (Figure 4E). This mechanism would explain how Aspirin initially acts as a substrate for CAII esterase activity, then its product, SA, is able to inhibit the enzyme."

"A typical dose of Aspirin is 325 mg; however, there are lower dosage options for everyday use and higher concentrations (up to 6 g per day, or ~7 mM in blood) for at-risk patients with heart disease [22]."

"At physiologically relevant concentrations of SA, it was shown that SA completely inhibited CAII (Figure 2). Using prism 8 software, the data was plotted to a non-linear regression to determine IC50 [41]. The average standard deviation was 2.2% and the standard deviation specifically at 50% activity is 1.5%. With the small deviation in percent activity, we can confidently say that the IC50 is 6.6 +/− 0.5 mM (Figure 2). Therefore, we show that at a clinically prescribed high dosage of Aspirin, CAII can act as an Aspirin esterase to form SA, which can then act as a suicide inhibitor."

"Based on these findings, we conclude that Aspirin binds and inhibits CAII via the SA product, as it retains the carboxylic acid motif similar to other CAIs such as nicotinic, ferulic, and 3-nitrobenzoic acids. These findings imply CAII’s importance in the blood, beyond its carbonic anhydrase activity and further implicate the enzyme in platelet function. We have identified CAII as the carboxylesterase responsible for Aspirin’s short half-life in the blood. Therefore, perhaps a combined therapy with a CA inhibitor and Aspirin could improve Aspirin’s efficacy in the treatment of heart disease."

 

[Dr. B]

I came across the following publication after our ??n's latest class.

- Aspirin: A Suicide Inhibitor of Carbonic Anhydrase II

Spoiler

"In addition to sulfonamides, a variety of other chemical motifs have been identified to inhibit CA, such as carboxylic acids [15]. Nicotinic and ferulic acid have recently been identified as inhibitors of CAII [16]. Unlike the sulfonamide-based drugs, these inhibitors do not directly displace the zinc bound solvent, but instead anchor through the solvent, blocking substrate entry to the active site [16]. Furthermore, 3-nitrobenzoic acid has also been reported as a potent CAI, with further studies showing its potential clinical relevance as a cancer therapeutic [17]. Previous work has also shown that salicylic acid as well as some phenol derivatives are μM inhibitors of mammalian CAs, although the exact mechanism of inhibition for salicylic acid is unknown [18]. These carboxylic acid-based compounds represent a new and largely unstudied class of CAIs."

Spoiler

​

"A recent study found in a genome-wide search that CAII is the only protein overexpressed in patients with Aspirin resistance and therefore may be the unidentified carboxylesterase [24]. Since Aspirin is a carboxylic acid-based molecule, it was hypothesized that it could potentially bind to CAII. Here, we examine this hypothesis through structure activity relationship studies between Aspirin and CAII, through X-ray crystallography and a spectroscopy-based kinetic assay. We determine that CAII is the previously unidentified carboxylesterase responsible for Aspirin’s short half-life in the blood, and that the product of this reaction, SA, can then inhibit CAII, thus making Aspirin a suicide inhibitor."​

​

"A known and widely studied function of CAs is their ability to act as an esterase [37]. When a small molecule with an ester bond such as Aspirin enters the active site of CAII, its ester bond is cleaved leaving an acetyl group and SA in the case of Aspirin. The zinc bound hydroxide is a strong nucleophile, able to attack the carbonyl of an ester, cleaving the bond."​

​

"Based on the crystallographic data of SA bound to CAII and the modeling with Aspirin, we propose a mechanism for CAII Aspirin ester cleavage and SA inhibition (Figure 4). Firstly, Aspirin binds to the zinc bound solvent within the active site with its acetate bound in a similar fashion of CO2 binding, positioned for nucleophilic attack (Figure 4A). The hydroxyl cleaves the ester bond in the Aspirin molecule leaving the acetate bound to the active site (Figure 4B). The acetate of the reaction is displaced by a water molecule that binds the zinc, while the SA remains in the active site (Figure 4C,D). Finally, the SA reorients within the active site and anchors through the zinc bound solvent, inhibiting any further reaction (Figure 4E). This mechanism would explain how Aspirin initially acts as a substrate for CAII esterase activity, then its product, SA, is able to inhibit the enzyme."​

​

"A typical dose of Aspirin is 325 mg; however, there are lower dosage options for everyday use and higher concentrations (up to 6 g per day, or ~7 mM in blood) for at-risk patients with heart disease [22]."​

​

"At physiologically relevant concentrations of SA, it was shown that SA completely inhibited CAII (Figure 2). Using prism 8 software, the data was plotted to a non-linear regression to determine IC50 [41]. The average standard deviation was 2.2% and the standard deviation specifically at 50% activity is 1.5%. With the small deviation in percent activity, we can confidently say that the IC50 is 6.6 +/− 0.5 mM (Figure 2). Therefore, we show that at a clinically prescribed high dosage of Aspirin, CAII can act as an Aspirin esterase to form SA, which can then act as a suicide inhibitor."​

​

"Based on these findings, we conclude that Aspirin binds and inhibits CAII via the SA product, as it retains the carboxylic acid motif similar to other CAIs such as nicotinic, ferulic, and 3-nitrobenzoic acids. These findings imply CAII’s importance in the blood, beyond its carbonic anhydrase activity and further implicate the enzyme in platelet function. We have identified CAII as the carboxylesterase responsible for Aspirin’s short half-life in the blood. Therefore, perhaps a combined therapy with a CA inhibitor and Aspirin could improve Aspirin’s efficacy in the treatment of heart disease."​

​

what about caffeine, rosaminic acid, pomegranate juice, green tea catechins, how effective are those compared to thiamine mate!

 

[Amazoniac]

what about caffeine, rosaminic acid, pomegranate juice, green tea catechins, how effective are those compared to thiamine mate!

In spite of the mismatch in acetazolamine values, they can serve as reference to compare how others fare in relation.

- Thiamine Is A Carbonic Anhydrase Inhibitor As Effective As Acetazolamide

3 is thiamide.​

- Carbonic Anhydrase Is A Key Driver Of Aging; Inhibiting It Is Beneficial

- Haemodialysis in massive caffeine intoxication

"The consumption of ~0.5 mol (100 g) of pure caffeine is equivalent to ~1200 cups of regular coffee and resulted in a caffeine serum concentration of 2.95 mmol/L (574 mg/L). It is the highest caffeine overdose which has reportedly been survived to date. By contrast, in another reported case, the ingestion of 10 g caffeine proved fatal [4]. Peak caffeine serum concentrations after drinking three cups of coffee (1.29 mmol, 250 mg caffeine) are in the range of 0.04 [mM] (7 mg/L) [5]."​

 

[Dr. B]

In spite of the mismatch in acetazolamine values, they can serve as reference to compare how others fare in relation.

- Thiamine Is A Carbonic Anhydrase Inhibitor As Effective As Acetazolamide

View attachment 27260​

3 is thiamide.​

- Carbonic Anhydrase Is A Key Driver Of Aging; Inhibiting It Is Beneficial

View attachment 27261​

- Haemodialysis in massive caffeine intoxication

"The consumption of ~0.5 mol (100 g) of pure caffeine is equivalent to ~1200 cups of regular coffee and resulted in a caffeine serum concentration of 2.95 mmol/L (574 mg/L). It is the highest caffeine overdose which has reportedly been survived to date. By contrast, in another reported case, the ingestion of 10 g caffeine proved fatal [4]. Peak caffeine serum concentrations after drinking three cups of coffee (1.29 mmol, 250 mg caffeine) are in the range of 0.04 [mM] (7 mg/L) [5]."​

are these equivalent values, like 1mg caffeine = 1mg acetazolamide in those tables?
i wonder how the more natural sources like pomegranate juice, green tea catechins and rosaminic acid compare... i wonder if shilajit has any effects on carbonic anhydrase

 

[Amazoniac]

are these equivalent values, like 1mg caffeine = 1mg acetazolamide in those tables?
i wonder how the more natural sources like pomegranate juice, green tea catechins and rosaminic acid compare... i wonder if shilajit has any effects on carbonic anhydrase

No, they're in moles for a fair comparison. Converting to mass would account for differences between molecules.

 

[aliml]

Acipimox, a nicotinic acid derivative in clinical use for the treatment of hyperlipidaemia, inhibits human carbonic anhydrases

 

[Beastmode]

@haidut

Do you think the methods of increasing CO2 have different impacts depending on what you want to heal/fix? (i.e- bag breathing vs CO2 in a garbage that you sit in, etc)

 

[Bart1]

could methylene blue be regarded as an carbonic anhydrase inhibitor ?

 

[ddjd]

I mean eat the chamomile buds. I buy organic buds for tea but also eat the buds after brewing them, to get all the Apigenin that didn’t go into solution and the many other beneficial substances in it. It’s eccentric I know, but my wife and kids are used to it. And I‘m a healthy bastard so I and justify any of these eccentricities with „he who cures/is in health is right“.
You can just as well eat raw chamomile, it tastes floral-spicy. An interesting taste, put it into a salad

So eat the fresh chamomile or dried better??

Like the idea of eating the buds after brewing, I'll try that thanks