Biomimetic synergistic site probe and application thereof

Abstract

The application relates to a kind of bionic synergic site probes and its application, the structural general formula of the probe is as shown in formula I: I.The application significantly improves the recognition ability of sensor to ultra-low concentration fentanyl, ketamine and other opioid molecules by bionic synergic active site design, realizes specific capture and signal reading out;The sensor preparation method provided is simple, low in cost, suitable for portable, rapid on-site screening, and has wide application prospect in the field of public safety, drug enforcement and environmental monitoring.

Description

Technical Field

[0001] This invention belongs to the field of gas chemical sensing technology, and specifically relates to a biomimetic synergistic site probe and its application. Background Technology

[0002] Opioids are a class of high-risk drugs that act on the central nervous system, including synthetic opioids such as fentanyl and psychoactive substances such as ketamine. Related studies have shown that these substances are highly addictive, have a high latency period, and have low lethal doses. Long-term abuse can cause serious damage to the respiratory, nervous, and cardiovascular systems; even trace exposure can trigger significant acute poisoning reactions. Therefore, rapid, portable, and reliable detection of opioids is of significant practical importance in fields such as public safety and medical surveillance.

[0003] Currently, the detection of opioids both domestically and internationally mainly relies on the following technical methods:

[0004] (1) Chromatography-mass spectrometry (GC-MS, LC-MS): High sensitivity and good accuracy, but the instruments are expensive, bulky, and require professional operators, making it difficult to meet the needs of on-site testing;

[0005] (2) Immunoassay (test strip, antibody method): Although the operation is simple, it relies on the recognition of specific antigens and antibodies, and the cross-reactivity with novel structural analogs is insufficient, which easily leads to false negatives or false positives;

[0006] (3) Ion mobility spectrometry (IMS): It is applicable to some aerosols and volatiles, but it is easily affected by humidity and background interference, and its ability to distinguish compounds with similar structures is limited.

[0007] (4) Spectroscopic methods (Raman, infrared, etc.): can achieve non-destructive detection, but for opioids with low vapor pressure and low concentration, the signal is often weak and the background noise is high, making it difficult to achieve trace detection.

[0008] All of the aforementioned technologies rely on contact sampling, which cannot meet the needs of non-contact detection in practical applications. A crucial technical problem urgently needs to be solved: how to stably, reliably, and rapidly detect extremely low concentrations of opioid vapors emitted naturally at room temperature without the aid of solution or solid samples.

[0009] Nanomaterial-based resistive sensors have attracted much attention due to their fast response speed and ease of miniaturization and integration. However, current technologies still face bottlenecks. For example, pure carbon nanotube gas sensors are susceptible to interference from volatile organic compounds in the environment. Although some studies have attempted to modify carbon nanotubes through methods such as nitrogen doping, their detection limits (e.g., 1 ppm) and selectivity are still far from meeting the needs of practical trace gas detection. Other reports have described the use of organic polymers to construct field-effect transistor sensors, which, while improving the detection limit, suffer from problems such as incomplete recovery of the sensing curve and severe baseline drift, limiting their practical applications.

[0010] The precise drug recognition capabilities of bioreceptor proteins can provide a novel perspective for the design of sensitive sites. For example, the μ-opioid receptor, as a target of opioid drugs, has a key site consisting of a triangular stereosynthetic site formed by the hydrophobic region of aromatic residues (phenylalanine / tyrosine / tryptophan) and the hydrogen-bonded region (His297 imidazole group and Asp147 carboxyl group). The amino acid residues constituting the site are all typical organic groups, highly adapted to organic molecular building blocks and synthetic processes, making it suitable for constructing core sensitive structures based on drug receptor sites.

[0011] Therefore, by referring to the biological receptor recognition sites and recognition mechanisms, we can realize the customized design of highly selective and highly sensitive sensing probes and effectively integrate them into electrical sensor devices, which provides a good opportunity to solve the urgent drug detection problem in this field. Summary of the Invention

[0012] This invention provides a biomimetic synergistic site probe and its application to solve the problems of insufficient sensitivity, poor selectivity, lack of portable equipment, and detection limit that cannot meet practical needs in the existing technology for rapid, portable, and reliable detection of opioids.

[0013] This invention provides a biomimetic cooperative site probe, the general structural formula of which is shown in Formula I and Figure 1 As shown:

[0014] I;

[0015] Wherein, A1 includes one or more of the following: benzene ring, naphthalene ring, anthracene ring, pyrene ring, thiophene ring, furan ring, pyrrole ring, carbazole group, fluorene group, and benzothiadiazole group;

[0016] The A2 comprises one or more of the following: benzene ring, naphthalene ring, anthracene ring, pyrene ring, thiophene ring, furan ring, pyrrole ring, carbazole group, fluorene group, and benzothiadiazole group, wherein X and R1 are substituted thereon;

[0017] X is a group containing N, O, S, P, F, Cl, Br or I that is inherent in the molecular skeleton or introduced through a side chain;

[0018] R1 includes one or more of hydrogen atoms, alkyl chains, alkoxy chains, ester chains, amide chains, fluorophores, and halogen substituents. The probe consists of an aromatic ring host structure with fluorescence emission properties, and at least two aromatic sites containing side chains and one heteroatom substitution site, which together form a cooperative active site with a characteristic spatial arrangement.

[0019] Preferably, the probe has the following structural formula: Figure 3 As shown.

[0020] The present invention also provides a method for preparing the aforementioned biomimetic synergistic site probe, comprising the following steps:

[0021] Under the combined action of palladium catalyst, base, and high-boiling-point organic solvent, and through heating in an inert atmosphere, an aromatic heterocyclic skeleton containing halogen substituents is coupled with a boric acid / boronic acid ester derivative having aromatic or heteroatom sites via Suzuki coupling to obtain the biomimetic synergistic site probe, such as... Figure 2 As shown.

[0022] The present invention also provides a sensing material comprising the aforementioned biomimetic synergistic site probe and carrier material, wherein the biomimetic synergistic site probe and carrier material form a sensing interface through non-covalent interaction.

[0023] Preferably, the carrier material includes one or more of the following: carbon nanotubes, graphene, reduced graphene oxide, carbon nanofibers, metal oxide nanowires (such as ZnO, SnO2), conductive polymers (such as polyaniline, polypyrrole), metal-organic frameworks, and covalent organic frameworks.

[0024] The present invention also provides a sensor comprising the aforementioned sensing material. This sensor is typically a resistive sensor or a field-effect transistor sensor, and detects opioid molecules by monitoring changes in the resistance or current / voltage of the sensing material before and after exposure to a target gas.

[0025] The present invention also provides a method for manufacturing the aforementioned sensor, comprising the following steps:

[0026] The biomimetic synergistic site probe and the carrier material are mixed in a solvent and a uniform dispersion is formed by means of ultrasound, which is the precursor of the sensing material. Then, the precursor liquid is coated onto a substrate with electrodes by means of dip coating, spin coating, drop coating or screen printing. After the solvent evaporates and dries, the sensor can be obtained.

[0027] Preferably, the solvent is selected from one or more of N,N-dimethylformamide, dimethyl sulfoxide, chloroform, tetrahydrofuran, carbon tetrachloride, acetonitrile, toluene, chlorobenzene, benzene, and dichloromethane.

[0028] Preferably, the electrodes include, but are not limited to, interdigitated electrodes and fractal electrodes.

[0029] This method is simple to operate, low in cost, and easy to implement for large-scale production and device integration.

[0030] This invention also provides an application of the aforementioned sensing material or sensor in the identification, detection, and monitoring of opioid drug molecules. For example, when the sensor is placed in an atmosphere containing the gas, the sensor's resistance changes, thus enabling the detection of opioid drug molecule gases.

[0031] Preferably, the opioid drug molecule includes one of fentanyl, ketamine, morphine, heroin, OxyContin, memantine, naloxone, and other opioid antagonists.

[0032] Preferably, the lower limit of sensor detection is 1ppt.

[0033] Based on the highly selective recognition of biomimetic synergistic sites, the sensor of this invention can produce a specific response to trace opioid drug molecules in complex environments (such as air, urine, blood, sewage, etc.), enabling rapid, portable, and sensitive on-site screening. The detection limit can reach the ppt or even ppq level, and it has significant application value and broad market prospects in the fields of public safety, drug enforcement, clinical diagnosis, and environmental monitoring.

[0034] Beneficial effects

[0035] This invention significantly improves the sensor's ability to identify ultra-low concentrations of opioid drug molecules such as fentanyl and ketamine through biomimetic synergistic active site design, achieving specific capture and signal readout. The sensor preparation method provided is simple and inexpensive, suitable for portable and rapid on-site screening, and has broad application prospects in the fields of public safety, drug enforcement, and environmental monitoring. Attached Figure Description

[0036] Figure 1 This is the general structural formula for the biomimetic collaborative site probe in this invention.

[0037] Figure 2 This is the general synthesis process for the biomimetic synergistic site probe in this invention.

[0038] Figure 3 These are the chemical structural formulas of the two biomimetic synergistic site probes prepared in Example 1.

[0039] Figure 4 This describes the synthesis process of the two biomimetic synergistic site probes prepared in Example 1.

[0040] Figure 5 These are scanning electron microscope images of the two biomimetic synergistic site probes prepared in Example 1.

[0041] Figure 6 These are scanning electron microscope images of the two biomimetic synergistic site probes prepared in Example 1 after being attached to short single-walled carbon nanotubes.

[0042] Figure 7 This is a schematic diagram of the test apparatus in Example 3.

[0043] Figure 8 The figures show the resistance values ​​of the two biomimetic synergistic site probes prepared in Example 1 against different concentrations of opioid drug molecule gas.

[0044] Figure 9a -b is a graph showing the response values ​​of the two biomimetic synergistic site probes prepared in Example 1 to common interfering gases.

[0045] Figure 10a -b is the 1H NMR spectrum of the two biomimetic synergistic site probes prepared in Example 1. Detailed Implementation

[0046] The present invention will be further illustrated below with reference to specific embodiments. It should be understood that these embodiments are for illustrative purposes only and are not intended to limit the scope of the invention. Furthermore, it should be understood that after reading the teachings of this invention, those skilled in the art can make various alterations or modifications to the invention, and these equivalent forms also fall within the scope defined by the appended claims.

[0047] For the terms defined below, unless a different definition is given elsewhere in the claims or this specification, these definitions shall apply. All numerical values, whether explicitly indicated or not, are defined herein as being modified by the term "about." The term "about" generally refers to a range of numerical values ​​that a person skilled in the art would consider equivalent to the stated values ​​to produce substantially the same properties, functions, results, etc. A range of numerical values ​​indicated by a low value and a high value is defined as including all numerical values ​​included within that range and all subranges included within that range.

[0048] It should be noted that the terms "comprising" and "having," and any variations thereof, in the specification, claims, and accompanying drawings of this disclosure are intended to cover non-exclusive inclusion.

[0049] Example 1

[0050] This embodiment provides a biomimetic synergistic site probe. In this embodiment, compounds containing carbazole or 2,1,3-benzothiadiazole are used as raw materials and synthesized with monomers containing 2-chloro-4-isopropyloxybenzene to obtain the final probe.

[0051] like Figure 4As shown, the following is the specific reaction process.

[0052] ① Under argon protection, 3,6-dibromocarbazole (325 mg, 1 mmol), 2-chloro-4-isopropyloxyphenylboronic acid (857.8 mg, 4 mmol), potassium carbonate (830 mg, 6 mmol), [1,1'-bis(diphenylphosphine)ferrocene]palladium dichloromethane complex (395 mg, 4 mmol), and 1,4-dioxane (40 mL) were added to a 100 mL reaction flask. The mixture was heated to 90 °C and reacted for 48 hours. The solution was then poured into a 100 mL ice-water mixture and extracted three times with dichloromethane (150 mL). The organic phase was dried over anhydrous magnesium sulfate and then evaporated to dryness. The solution was then passed through a column to obtain a white solid of 302.4 mg, with a yield of 60%, which was identified as probe 1.

[0053] ② Under argon protection, 3,6-dibromocarbazole (325 mg, 1 mmol), 2-chloro-4-isopropoxyphenylboronic acid pinacol ester (1.186 g, 4 mmol), potassium carbonate (830 mg, 6 mmol), [1,1'-bis(diphenylphosphine)ferrocene]palladium dichloride dichloromethane complex (395 mg, 4 mmol), and 1,4-dioxane (40 mL) were added to a 100 mL reaction flask. The mixture was heated to 90 °C and reacted for 48 hours. The solution was poured into a 100 mL ice-water mixture and extracted three times with dichloromethane (150 mL). The organic phase was dried over anhydrous magnesium sulfate and then evaporated to dryness. The solution was passed through a column to obtain a white solid of 354.8 mg, with a yield of 70%, which was probe 1.

[0054] The proton NMR spectrum of probe 1 is shown below. Figure 10a As shown.

[0055] ③ Under argon protection, 4,7-dibromo-2,1,3-benzothiadiazole (294 mg, 1 mmol), 2-chloro-4-isopropyloxyphenylboronic acid (857.8 mg, 4 mmol), potassium carbonate (830 mg, 6 mmol), [1,1'-bis(diphenylphosphine)ferrocene]palladium dichloride dichloromethane complex (395 mg, 4 mmol), and 1,4-dioxane (40 mL) were added to a 100 mL reaction flask. The mixture was heated to 90 °C and reacted for 48 hours. The solution was poured into a 100 mL ice-water mixture and extracted three times with dichloromethane (150 mL). The organic phase was dried over anhydrous magnesium sulfate and then evaporated to dryness. The solution was then passed through a column to obtain a white solid of 378.4 mg, with a yield of 80%, which was probe 2.

[0056] ④ Under argon protection, 4,7-dibromo-2,1,3-benzothiadiazole (294 mg, 1 mmol), pinacol ester of 2-chloro-4-isopropoxyphenylboronic acid (1.186 g, 4 mmol), potassium carbonate (830 mg, 6 mmol), [1,1'-bis(diphenylphosphine)ferrocene]palladium dichloride dichloromethane complex (395 mg, 4 mmol), and 40 mL of 1,4-dioxane were added to a 100 mL reaction flask. The mixture was heated to 90 °C and reacted for 48 hours. The solution was poured into a 100 mL ice-water mixture and extracted three times with dichloromethane (150 mL). The organic phase was dried over anhydrous magnesium sulfate and then evaporated to dryness. The solution was passed through a column to obtain a white solid of 236.5 mg, with a yield of 50%, which was probe 2.

[0057] The proton NMR spectrum of probe 2 is shown below. Figure 10b As shown.

[0058] Example 2

[0059] Using 10 mL of N,N-dimethylformamide as solvent, 0.4 mg of biomimetic synergistic site probe 1 and 0.2 mg of short single-walled carbon nanotubes were mixed, with a probe-to-carbon nanotube material mass ratio of 2:1. The mixture was ultrasonically mixed for 10 min to obtain a homogeneous solution. The scanning electron microscope image of the pure sensing material is shown below. Figure 5 As shown in the image, the scanning electron microscope image of the sensing material with attached carbon nanotubes is as follows: Figure 6 As shown, it can be seen that after carbon nanotubes and sensing probes are attached through non-covalent interactions, the materials are more uniformly dispersed, which is beneficial to improving the sensing effect.

[0060] 10 mL of N,N-dimethylformamide was used as the solvent, along with 2 mg of biomimetic synergistic site probe 2 and 0.2 mg of short single-walled carbon nanotubes, with a mass ratio of probe to carbon nanotube material of 10:1. The mixture was ultrasonically mixed for 10 min to obtain a homogeneous solution. A scanning electron microscope image of the pure sensing material is shown below. Figure 5 As shown in the image, the scanning electron microscope image of the sensing material with attached carbon nanotubes is as follows: Figure 6 As shown, it can be seen that after carbon nanotubes and sensing probes are attached through non-covalent interactions, the materials are more uniformly dispersed, which is beneficial to improving the sensing effect.

[0061] Example 3

[0062] First, an interdigitated electrode was used as the test electrode. A 1 μL pipette was used to coat the test electrode with the mixture from Example 2. After the solvent evaporated, a sensor based on a biomimetic sensitive structure was obtained. A schematic diagram of the test setup is shown below. Figure 7 As shown.

[0063] At 25°C in a sealed vacuum environment, a series of test gases of different concentrations were prepared by mixing standard opioid gases, such as fentanyl and ketamine, with nitrogen. The volume fractions of fentanyl were 100 ppt, 50 ppt, 25 ppt, 20 ppt, 10 ppt, 2 ppt, and 1 ppt. The volume fractions of ketamine were 700 ppt, 350 ppt, 200 ppt, 175 ppt, 100 ppt, 5 ppt, and 1 ppt. The sensor was placed inside the cavity, and the device was connected to the circuit and a resistance acquisition device to monitor the resistance changes. Sensitivity (response intensity) S = ΔR / R0 (S is the sensitivity, R0 is the resistance at the start time, and ΔR is the resistance at the end time minus the resistance at the start time). By continuously introducing approximately 50 mL of a certain concentration of the gas to be measured, a significant decrease in resistance can be observed. After the resistance recovers, an even lower concentration of the gas to be measured is introduced, and this process is repeated to obtain a sensing curve showing the change in resistance over time. Figure 8 As shown, this sensor can effectively sense opioid gases.

[0064] Example 4

[0065] Considering that in actual testing scenarios, sensing opioid drug molecules in the atmospheric environment would be subject to interference from various non-opioid drug molecules, such as ethyl acetate, aniline, toluene, chlorobenzene, acetic acid, methanol, ethanol, water vapor, air, tetrahydrofuran, and other common solvents, as well as other non-opioid drugs like MPEA, gas-phase sensing tests were performed on the interfering gases using the same method as in Example 3. The interfering gases were: 10,000 ppm ethyl acetate, 100 ppb aniline, 2,500 ppm toluene, 1,300 ppm chlorobenzene, 200 ppm acetic acid, 100,000 ppm methanol, 7,000 ppm ethanol, 32 ppm water vapor, air, 20,000 ppm tetrahydrofuran, and 3 ppb MPEA. As shown in Figure 9, it can be seen that the sensor has strong anti-interference capabilities and good selectivity against these non-opioid drug molecules.

Claims

1. A biomimetic cooperative site probe, characterized in that, The general structural formula of the probe is shown in Formula I: I； Wherein, A1 includes one or more of the following: benzene ring, naphthalene ring, anthracene ring, pyrene ring, thiophene ring, furan ring, pyrrole ring, carbazole group, fluorene group, and benzothiadiazole group; The A2 comprises one or more of the following: benzene ring, naphthalene ring, anthracene ring, pyrene ring, thiophene ring, furan ring, pyrrole ring, carbazole group, fluorene group, and benzothiadiazole group, wherein X and R1 are substituted thereon; X is a group containing N, O, S, P, F, Cl, Br or I that is inherent in the molecular skeleton or introduced through a side chain; R1 includes one or more of the following: hydrogen atom, alkyl chain, alkoxy chain, ester chain, amide chain, fluorophore, and halogen substituent.

2. A method for preparing the biomimetic synergistic site probe as described in claim 1, characterized in that, Includes the following steps: The biomimetic synergistic site probe is obtained by heating the reaction under an inert atmosphere in the presence of a palladium catalyst, a base, and a high-boiling-point organic solvent, and then coupling the aromatic heterocyclic skeleton containing halogen substituents with boric acid / boronic acid ester derivatives with aromatic or heteroatom sites via Suzuki coupling.

3. A sensing material, characterized in that, It includes the biomimetic synergistic site probe and carrier material as described in claim 1, wherein the biomimetic synergistic site probe and carrier material form a sensing interface through non-covalent interaction.

4. The sensing material according to claim 3, characterized in that, The carrier material includes one or more of the following: carbon nanotubes, graphene, reduced graphene oxide, carbon nanofibers, metal oxide nanowires, conductive polymers, metal-organic frameworks, and covalent organic frameworks.

5. A sensor, characterized in that, It includes the sensing material as described in claim 3.

6. A method for manufacturing a sensor as described in claim 5, characterized in that, Includes the following steps: The biomimetic synergistic site probe is mixed with a carrier material in a solvent to form a sensing material; the sensing material is coated onto a substrate with electrodes and dried to obtain the sensor.

7. The application of a sensing material as described in claim 3 or a sensor as described in claim 5 in the identification, detection, and monitoring of opioid drug molecules.

8. The application according to claim 7, characterized in that, The opioid drug molecules include one of the following: fentanyl, ketamine, morphine, heroin, OxyContin, memantine, naloxone, and other opioid antagonists.

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