Electron donor-acceptor-donor type ruthenium complex as well as preparation method and application thereof

Abstract

The invention discloses an electron donor-acceptor-donor type ruthenium complex as well as a preparation method and application thereof. The structural formula of the ruthenium complex is as follows: # imgabs0 #, wherein X <-> represents an anion. The ruthenium complex disclosed by the invention has a relatively strong photodynamic treatment effect on a human non-small cell lung cancer cell line (A549 cells). The complex can be excited only by red light illumination, and has higher compatibility with existing medical equipment; meanwhile, the light penetrability of red light is weaker than that of near-infrared light, so that the red light is more suitable for superficial tumor treatment. Due to the moderate PI value, immunosuppression or tissue damage caused by excessive killing can be avoided, so that the compound can be suitable for multiple times of treatment or combined use with other therapies (for example, when the compound is combined with chemotherapy, immunotherapy or radiotherapy and the like, the compound can be used as an auxiliary treatment means, and active oxygen (ROS) is generated through photocatalysis to enhance the effects of other therapies).

Description

Technical Field

[0001] The present invention relates to the technical field of medicinal chemistry, and in particular to an electron donor-acceptor-donor type ruthenium complex and a preparation method and application thereof. Background Art

[0002] Lung cancer, also known as primary bronchogenic carcinoma, is a common lung malignancy originating from the trachea, bronchial mucosa, or glands. Based on its histopathological characteristics, lung cancer is mainly divided into two categories: non-small cell lung cancer (NSCLC) and small cell lung cancer (SCLC). Among them, non-small cell lung cancer includes multiple subtypes, including central type (such as squamous cell carcinoma) and peripheral type (such as adenocarcinoma), and has a significant clinical incidence rate. Therefore, the development of therapeutic drugs for non-small cell lung cancer has great medical value.

[0003] Due to its complex biological behavior and diverse clinical manifestations, non-small cell carcinoma (NSCLC) remains a research focus in the field of lung cancer treatment. In recent years, transition metal ruthenium complexes have demonstrated significant potential as photosensitizers (PS) in photodynamic therapy (PDT) due to their unique photophysical properties, including significant Stokes shifts, strong spin-orbit coupling, high quantum yield luminescence, long-lived phosphorescent states, and efficient triplet exciton generation. These innovative photosensitizers absorb light energy of specific wavelengths, triggering photochemical reactions within cells and inducing tumor cell apoptosis, opening up new avenues for the treatment of NSCLC.

[0004] CN116496323A discloses a binuclear ruthenium photocatalyst for photocatalytic oxidation of amino acids and its application. The compound has a strong ability to inhibit the growth and proliferation of cisplatin-resistant human non-small cell lung cancer cells under near-infrared light conditions. However, the compound is mainly suitable for the treatment of deep tumors (central type) and is difficult to use for the treatment of superficial tumors (peripheral type). At the same time, the compound needs to rely on near-infrared light when used for photodynamic therapy, which has higher requirements for medical equipment and low compatibility with existing medical equipment.

[0005] Therefore, although transition metal ruthenium complexes have made breakthrough progress in the field of PDT, their application in the treatment of non-small cell carcinoma still faces multiple challenges. For example, non-small cell carcinoma is highly heterogeneous, and tumors from different patients have significant differences in gene expression, metabolic pathways, and immune microenvironment, which makes it difficult for a single treatment strategy to achieve ideal results. In addition, the metastasis and recurrence of non-small cell carcinoma also need to be addressed urgently. Even after comprehensive treatment such as surgical resection and chemotherapy, some patients will still experience recurrence or distant metastasis, leading to treatment failure.

[0006] Therefore, to further improve the treatment efficacy of NSCLC and reduce the risk of recurrence and metastasis, the continuous exploration of new treatment strategies and drugs is necessary. This includes developing new photosensitizers with greater efficacy and fewer side effects, researching targeted drugs targeting specific NSCLC targets, and exploring new treatment options such as immunotherapy and gene therapy. These efforts are expected to provide NSCLC patients with more effective treatment options and improve their survival rates and quality of life. Summary of the Invention

[0007] The present invention aims to solve at least one of the technical problems existing in the prior art. To this end, the present invention provides an electron donor-acceptor-donor ruthenium complex that can be used to treat superficial tumors and has high compatibility with medical equipment.

[0008] The present invention also provides a method for preparing the ruthenium complex.

[0009] The present invention also provides the application of the ruthenium complex.

[0010] According to one aspect of the present invention, a ruthenium complex is provided, and the structural formula of the ruthenium complex is as follows:

[0011]

[0012] Among them, X - Represents anions.

[0013] According to a preferred embodiment of the present invention, at least the following beneficial effects are achieved: the ruthenium complex of the present invention has a strong photodynamic therapy effect on human non-small cell lung cancer cell line (A549 cells). Under red light illumination, it has a strong ability to inhibit the growth and proliferation of human non-small cell lung cancer cells (IC 50 is 2.49μM), while under dark conditions, its cytotoxicity is greater than 100μM, and the phototherapy index PI is as high as 40.2. The complex only requires red light to be excited, and is more compatible with existing medical equipment; at the same time, the penetration ability of red light is weaker than that of near-infrared light, making it more suitable for the treatment of superficial tumors. A moderate PI value can avoid immunosuppression or tissue damage caused by excessive killing, making it suitable for multiple treatments or combined with other therapies (such as when used in combination with chemotherapy, immunotherapy or radiotherapy, it can be used as an auxiliary treatment to enhance the effects of other therapies by generating reactive oxygen species (ROS) through photocatalysis); a moderate IC 50 The high-potency value also enables the complex to find a better balance between safety and efficacy when used as a photosensitizer, avoiding the problem of increased toxicity of normal cells in the illuminated area (especially light leakage or non-targeted areas) that may be caused by overly potent drugs.

[0014] In addition, the complex is a mononuclear complex, which reduces the use of precious metals and saves economic costs compared to traditional binuclear ruthenium complexes.

[0015] In some preferred embodiments of the present invention, X - PF6 - PF6 anion - It can improve the solubility of ruthenium complexes and enhance their anti-tumor effects.

[0016] According to another aspect of the present invention, a method for preparing the above-mentioned complex is provided, comprising the following steps:

[0017] S1. 3+ 4'-Bromo-2,2':6',2"-terpyridine is mixed with a solvent and heated to obtain intermediate I;

[0018] S2. Add X to the reaction solution in step S1. - to obtain intermediate II;

[0019] S3. The intermediate II obtained in step S2 is reacted with 2,5-bis(2-ethylhexyl)-3,6-bis(thiophen-2-yl)pyrrolo[3,4-C]pyrrole-1,4(2H,5H)-dione to generate the ruthenium complex.

[0020] The preparation method according to a preferred embodiment of the present invention has at least the following beneficial effects: the preparation method of the present invention is easy to operate and has good industrial application prospects.

[0021] In some embodiments of the present invention, step S1 specifically comprises mixing ruthenium (III) chloride or its hydrate and 4'-bromo-2,2':6',2"-terpyridine, adding a solvent, and reacting under heating.

[0022] In some preferred embodiments of the present invention, Ru 3+ The molar ratio of 4'-bromo-2,2':6',2"-terpyridine is 1:2 to 2.2.

[0023] In some preferred embodiments of the present invention, Ru 3+ The molar ratio of 4'-bromo-2,2':6',2"-terpyridine is 1:2.1.

[0024] In some embodiments of the present invention, the solvent is selected from ethylene glycol.

[0025] In some embodiments of the present invention, in step S1, the reaction under heating includes at least one of the following conditions:

[0026] 1) The temperature is 180-200°C;

[0027] 2) The duration is 10 to 30 minutes;

[0028] 3) The heating method is heating reflux.

[0029] In some preferred embodiments of the present invention, in step S1, the reaction under heating includes at least one of the following conditions:

[0030] 1) Temperature is 200°C;

[0031] 2) The duration is 20 minutes;

[0032] 3) The heating method is heating reflux.

[0033] In some embodiments of the present invention, step S3 specifically includes reacting the ruthenium complex intermediate with a palladium catalyst (preferably palladium acetate), a carbonate (preferably potassium carbonate), pivalic acid and 2,5-bis(2-ethylhexyl)-3,6-bis(thiophen-2-yl)pyrrolo[3,4-C]pyrrole-1,4(2H,5H)-dione in an aprotic polar solvent to generate a target ruthenium complex.

[0034] In some embodiments of the present invention, the aprotic polar solvent is dimethylacetamide.

[0035] In some embodiments of the present invention, the step S3 is carried out under heating conditions.

[0036] In some embodiments of the present invention, the heating temperature in step S3 is selected from 90 to 110° C., and the heating time is selected from 4 to 8 hours.

[0037] In some preferred embodiments of the present invention, the heating temperature in step S3 is 110°C.

[0038] In some embodiments of the present invention, the molar ratio of the ruthenium complex intermediate to palladium acetate, potassium carbonate, pivalic acid and 2,5-bis(2-ethylhexyl)-3,6-bis(thiophen-2-yl)pyrrolo[3,4-C]pyrrole-1,4(2H,5H)-dione is 1:0.04~0.06:2~2.2:0.08~0.12:2~2.2.

[0039] In some preferred embodiments of the present invention, the molar ratio of the ruthenium complex intermediate to palladium acetate, potassium carbonate, pivalic acid and 2,5-bis(2-ethylhexyl)-3,6-bis(thiophen-2-yl)pyrrolo[3,4-C]pyrrole-1,4(2H,5H)-dione is 1:0.05:2:0.1:2.1.

[0040] According to another aspect of the present invention, the use of the above-mentioned complex in the preparation of anti-tumor drugs is proposed.

[0041] According to the application of a preferred embodiment of the present invention, there are at least the following beneficial effects: the complex of the present invention can generate superoxide anions under 635nm wavelength light. Superoxide anions are usually associated with oxidative stress in the body. They can participate in various physiological and pathological processes as signal molecules. Therefore, it can kill tumor cells or inhibit their growth by inducing oxidative stress. The complex of the present invention has a strong ability to generate superoxide anions after light irradiation, which can cause oxidative stress in tumor cells. In addition, the complex of the present invention can directly oxidize NADH through photocatalysis to generate NAD+ free radicals. This process does not rely on oxygen, so the complex is still effective in hypoxic tumors. In summary, it has good anti-tumor potential, which shows that it has good application prospects in the field of anti-tumor drugs.

[0042] In some embodiments of the present invention, the anti-tumor drug is an anti-non-small cell lung cancer drug. The complex of the present invention has a strong effect of inhibiting the proliferation of non-small cell lung cancer cells and has good application prospects in the treatment of non-small cell lung cancer.

[0043] In some embodiments of the present invention, the non-small cell lung cancer is A549 cell line lung cancer.

[0044] In some embodiments of the present invention, the anti-non-small cell lung cancer drug is a photodynamic therapy drug.

[0045] In some embodiments of the present invention, the wavelength of the photodynamic therapy drug excitation light is 400 to 640 nm.

[0046] In some embodiments of the present invention, the excitation light wavelength is 635 nm.

[0047] In some embodiments of the present invention, the drug is scientifically prepared by combining the active ingredient of the ruthenium complex with a pharmaceutical matrix. The matrix system contains functional carrier materials and pharmaceutical excipients, and is constructed into a dosage form that meets clinical needs through precise proportioning.

[0048] In some embodiments of the present invention, the formulation of the pharmaceutical matrix requires comprehensive consideration of several key factors: the use of inert fillers to ensure drug stability, the utilization of hydrophilic / hydrophobic balance modifiers to optimize bioavailability, the addition of disintegrants to control drug release characteristics, and the introduction of antioxidants to extend the shelf life of the formulation. New pharmaceutical excipients include polymers such as hydroxypropyl-β-cyclodextrin, chitosan microspheres, polyethylene glycol-polylactic acid copolymers, and sodium hyaluronate, as well as interface modification ingredients such as surfactants, suspending agents, and emulsifiers.

[0049] In some embodiments of the present invention, pharmaceutical formulation engineering achieves multi-objective optimization through the synergistic effects of pharmaceutical excipients: constructing nanoscale drug delivery systems to enhance targeting, regulating the solubility and membrane permeability of photosensitizers, and utilizing inclusion technology to improve drug stability. Pharmaceutical excipients can be tailored to the specific clinical route of administration: for example, intravenous injections require a sterile, pyrogen-free, isotonic solution system, oral formulations require disintegrants and flavoring agents, and transdermal drug delivery systems require polymeric film-forming materials.

[0050] In some embodiments of the present invention, pharmaceutical dosage form designs exhibit diverse features, including hard / soft capsules, compressed / coated tablets, lyophilized powders for injection, oral solutions, suspensions, syrups, orally disintegrating tablets, sublingual tablets, topical gels, rectal suppositories, and enemas. For special application scenarios, novel drug delivery systems can be constructed using liposome delivery systems, protein nanocarriers (carrierized / non-carrierized), organic / inorganic nanoparticles, nanoemulsions and microemulsions, nanocrystal preparations, and complex solvent systems.

[0051] In some embodiments of the present invention, clinical administration methods are flexibly selected based on the drug's characteristics: oral administration requires special consideration of gastrointestinal stability (e.g., enteric-coated tablet design), parenteral administration requires control of particle size distribution and surface charge, and topical administration can achieve sustained release using gels or patches. For photosensitive drugs, formulation design must also consider both light-protection storage and post-administration photoactivation conditions to ensure therapeutic safety and efficacy.

[0052] According to another aspect of the present invention, an anti-tumor metal photosensitizer is provided, wherein the active ingredient of the anti-tumor metal photosensitizer includes the above-mentioned ruthenium complex.

[0053] Additional aspects and advantages of the present invention will be set forth in part in the description which follows and, in part, will be obvious from the description which follows, or may be learned by practice of the present invention. BRIEF DESCRIPTION OF THE DRAWINGS

[0054] Figure 1 is the ultraviolet absorption spectrum of the metal ruthenium complex prepared in the embodiment of the present invention;

[0055] Figure 2 1 is a graph showing the test results of the photocatalytic oxidation ability of the ruthenium complex prepared in an embodiment of the present invention for NADH;

[0056] Figure 3 This is a graph showing the test results of the ability of the metal ruthenium complex prepared in an embodiment of the present invention to generate superoxide anions;

[0057] Figure 4 Graph showing the dark toxicity and phototoxicity test results of the ruthenium complex prepared in an embodiment of the present invention against human non-small cell lung cancer cells (A549). DETAILED DESCRIPTION

[0058] The following will clearly and completely describe the concept of the present invention and the technical effects produced in conjunction with the embodiments, so as to fully understand the purpose, characteristics and effects of the present invention. Obviously, the embodiments described are only some embodiments of the present invention, not all embodiments. Based on the embodiments of the present invention, other embodiments obtained by those skilled in the art without creative work are within the scope of protection of the present invention. Unless otherwise specified, the test methods used in the embodiments are all conventional methods; the materials, reagents, etc. used, unless otherwise specified, are reagents and materials obtained from commercial channels. Unless otherwise specified, the same parameter in each embodiment has the same value. The embodiments described below are exemplary and are only used to explain the present invention, and are not to be understood as limitations of the present invention.

[0059] In the description of the present invention, reference to the term "some embodiments" or the like indicates that a specific feature, structure, material, or characteristic described in conjunction with that embodiment or example is included in at least one embodiment or example of the present invention. In this specification, schematic representations of the above terms do not necessarily refer to the same embodiment or example. Furthermore, the specific features, structures, materials, or characteristics described may be combined in any suitable manner in any one or more embodiments or examples.

[0060] In the description of the present invention, if there is a description of first, second, etc., it is only for the purpose of distinguishing technical features, and cannot be understood as indicating or implying relative importance or implicitly indicating the number of the indicated technical features or implicitly indicating the order of the indicated technical features.

[0061] The "room temperature" referred to in the present invention refers to 25±5°C, and is specifically 25°C in the embodiments.

[0062] Example

[0063] In this example, a ruthenium complex was prepared. The complex can be excited by red light. Its structural formula is as follows:

[0064]

[0065] The specific preparation process is as follows:

[0066] S1. Ruthenium (III) chloride hydrate (0.208 g, 1 mmol) and 4'-bromo-2,2':6',2"-terpyridine (0.624 g, 2 mmol) were added to a reaction tube, and ethylene glycol (8 mL) was added. The mixture was then heated under reflux at 200°C for 20 minutes to obtain ruthenium complex intermediate I.

[0067] The chemical reaction equation above is as follows:

[0068]

[0069] S2. To the cooled reaction solution was added 10 ml of a saturated aqueous solution of ammonium hexafluorophosphate to precipitate the target ruthenium complex intermediate II. After cooling to room temperature, the red precipitate was filtered off and washed with ice water to obtain the ruthenium complex intermediate II with a yield of 70.4%.

[0070] S3. The ruthenium complex intermediate II (0.088 g, 0.1 mmol) obtained in step S2, palladium acetate (0.0013 g, 0.005 mmol), potassium carbonate (0.02 mmol), pivalic acid (0.001 g, 0.01 mmol) and 2,5-bis(2-ethylhexyl)-3,6-bis(thiophen-2-yl)pyrrolo[3,4-C]pyrrole-1,4(2H,5H)-dione (0.109 g, 0.21 mmol) were added to a reaction tube for argon protection and N,N-dimethylacetamide (3 mL) was added. The mixture was then heated under reflux at 110°C for 6 hours. After the reactant was cooled to room temperature, water (10 mL) was added and filtered to obtain a blue precipitate. The precipitate was washed with water, dried and purified to obtain the target ruthenium complex with a yield of 15.4%.

[0071] The chemical reaction equation above is as follows:

[0072]

[0073] The product obtained in the above step S3 was subjected to structural characterization, and the mass spectrum was as follows: ESI-MS [CH3OH, m / z]: 806.25 [M-2PF6 - ] 2+ .

[0074] The H NMR spectrum of the product is: 1 H NMR(500MHz,DMSO-d6)δ9.42(s,4H),9.11(d,J=8.1Hz,4H),8.88(d,J=3.9H z,4H),8.68–8.61(m,2H),8.18(d,J=4.9Hz,2H),8.12(d,J=7.7Hz,4H),7.6 4(d,J=5.7Hz,4H),7.45(s,2H),7.31(t,J=6.7Hz,4H),4.16(d,J=7.4Hz,4H ),4.04–4.02(m,5H),1.84–1.75(m,12H),1.29(s,24H),0.87–0.83(m,24H).

[0075] Test example

[0076] In order to verify the application effect of the above complex, its performance was tested, as follows:

[0077] 1. Absorption spectrum determination of ruthenium complex

[0078] The ruthenium complex prepared by the above operation was prepared into a 10 μM sample solution using phosphate-balanced saline (PBS) and acetonitrile (MeCN) as solvents, respectively. Then, a double-beam UV-visible spectrophotometer was used to record the UV absorption spectrum of the new red-light-excited ruthenium complex to characterize its absorbance in phosphate-balanced saline and acetonitrile. The results are as follows: Figure 1 shown.

[0079] from Figure 1 It can be seen from the figure that the complex prepared in the embodiment of the present invention has good light absorption ability in organic solvents.

[0080] 2. Determination of the ability of ruthenium complexes to photocatalytically oxidize NADH

[0081] Under light irradiation, the metal complex can oxidize the reduced coenzyme I (NADH) into its oxidized form NAD + Therefore, the ruthenium complex (10 μM) and NADH (A 339 nm≈1.0) in a cuvette and its ability to oxidize NADH under light conditions was measured. Figure 2 shown.

[0082] As can be seen from the figure, the ruthenium complex exhibits significant photocatalytic oxidation of NADH. This result demonstrates that the complex of the present invention can directly photocatalytically oxidize NADH to generate NAD+ free radicals. This process is independent of oxygen, demonstrating that the complex has oxygen-independent and multi-mechanistic anti-tumor potential, expanding its therapeutic application.

[0083] 3. Determination of the ability of ruthenium complexes to generate superoxide anions

[0084] The superoxide anion probe dihydrorhodamine 123 (DHR123) was used to determine the ability of the metal ruthenium complex to generate superoxide anions. The changes in the fluorescence intensity of the mixed solution of the test sample and DHR123 under different illumination times were monitored by a Techcomp FL970 fluorescence spectrophotometer to reflect the ability to generate superoxide anions.

[0085] A mixed solution of a ruthenium complex (10 μM) and DHR123 was excited in a 1 cm quartz tube at λex = 465 nm. The entrance and exit slits were set to 2.5 nm. The superoxide anion generation capacity was measured under 635 nm illumination. Figure 3It is shown that the metal ruthenium complex has the ability to generate superoxide anions after light irradiation.

[0086] 4. Photodynamic therapy effect of ruthenium complexes on human non-small cell lung cancer cell lines

[0087] Resazurin solution is blue and is commonly used as an acid-base indicator (orange at pH 3.8 to deep purple at pH 6.5) and a redox indicator. During cell viability assays, resazurin penetrates cells and is irreversibly reduced by living cells to a pink color, accompanied by the appearance of red fluorescent resorufin. The absorbance or fluorescence intensity of resorufin is positively correlated with cell number and reducing capacity, allowing analysis of cell proliferation using an enzyme-linked immunosorbent assay (ELISA).

[0088] The resazurin experimental steps are as follows:

[0089] (1) First, revive one tube of A549 tumor cells and culture them with fresh complete culture medium (DMEM medium + 10 vol% fetal bovine serum + 1 vol% penicillin-streptomycin mixture). After passage twice, start the experiment.

[0090] (2) When the cells reached the logarithmic growth phase, they were seeded into two 96-well plates at a density of 5000 cells / well (100 μL of culture medium was used to culture cells in each well, one plate was for the light group and the other plate was for the dark control group), and cultured in a 37°C, 5% CO2 incubator.

[0091] (3) After the cells adhere to the wall, the original culture medium was aspirated and 100 μL of ruthenium complexes at six concentrations, 100, 50, 10, 1, 0.1, and 0.01 μM, were added to each well. The cells were gently shaken and incubated in a carbon dioxide incubator (37°C, 5% CO2) in the dark.

[0092] (4) After incubation for 6 h, the cell culture plates in the illumination group were placed under a 635 nm light source for 45 min (light dose of 63.7 J / cm 2 ), and then returned to the incubator for further incubation in the dark for 42 h (cells in the dark control group were kept in the incubator in the dark for incubation).

[0093] (5) After incubation for 42 h, the culture medium was discarded from each well, and 80 μL of resazurin (100 mg / mL) was added to each well. The cells were incubated in a 37°C incubator for another 4 h. EX540 / EM590 was detected using the fluorescence plate of an enzyme-linked immunosorbent assay (ELISA) to calculate the cell proliferation inhibition rate and obtain the IC 50 value (drug concentration when the inhibition rate is equal to 50%).

[0094] The results are as follows Figure 4As shown in the figure, the resazurin method was used to detect the killing effect of different concentrations of ruthenium complexes on human non-small cell lung cancer cell line (A549 cells) under dark and light treatment conditions. It can be seen that in the absence of light, the IC 50 Greater than 100 μM, IC against human non-small cell lung cancer cell lines under light conditions 50 The phototherapy index PI is as high as 40.2, indicating that the metal ruthenium complex of the present invention has a strong photodynamic therapy effect.

[0095] The embodiments of the present invention are described in detail above, but the present invention is not limited to the above embodiments. Various changes can be made within the knowledge scope of ordinary technicians in the relevant technical field without departing from the scope of the present invention.

Claims

1. A ruthenium complex, characterized in that: The structural formula of the ruthenium complex is as follows: Among them, X - Represents anions.

2. The ruthenium complex according to claim 1, characterized in that: In the complex, X - PF6 - .

3. The method for preparing the complex according to claim 1 or 2, wherein: The steps include: S1. 3+ 4'-Bromo-2,2':6',2"-terpyridine is mixed with a solvent and heated to obtain intermediate I; S2. Add X to the reaction solution in step S1. - to obtain intermediate II; S3. The intermediate II obtained in step S2 is reacted with 2,5-bis(2-ethylhexyl)-3,6-bis(thiophen-2-yl)pyrrolo[3,4-C]pyrrole-1,4(2H,5H)-dione to generate the ruthenium complex.

4. Use of the complex according to claim 1 or 2 in the preparation of antitumor drugs.

5. The use according to claim 4, characterized in that: The anti-tumor drug is an anti-non-small cell lung cancer drug.

6. The use according to claim 5, characterized in that: The non-small cell lung cancer is A549 cell line lung cancer.

7. The use according to claim 5, characterized in that: The anti-non-small cell lung cancer drug is a photodynamic therapy drug.

8. The use according to claim 7, characterized in that: The wavelength of the drug excitation light for the photodynamic therapy is 400 to 640 nm.

9. The use according to claim 8, characterized in that: The excitation light wavelength is 635 nm.

10. An anti-tumor metal photosensitizer, characterized in that: The active ingredient of the anti-tumor metal photosensitizer includes the ruthenium complex as claimed in claim 1 or 2.

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